Abiogenesis: The Faith and the Facts

I'll be talking about abiogenesis tonight and that's one of those funny words that you can figure out what it means if you take it apart it currently it's the the most prevalent scientific explanation for the origin of life and when you when you take this word apart a is means not by all means life or living and Genesis is origin beginning or origin of life so this word is really selling us this is the non-biological origin of life and that's the current scientific explanation of how life arose that there was this non-biological process the origin life is one of those big questions in life it tests our faith taxes our science it challenges the deep thinkers it defines our identity and is an integral part of our worldview you can imagine that depending on how you imagine where life came from you're going to have different perspectives on what the human race is all about if we're just highly evolved pond scum you're going to have one view if you see yourself as a child of God you're going to have a totally different view and so this is one of those questions that is almost essential to civilization to get right and all civilizations one way or another try and develop answers to the question of where life came from tonight we're looking at the scientific aspects of this question so we want to get a definition of science that we all sort of agree upon so we'll have some common ground one definition of science is that it's a search for natural explanations of the world around us another one is that it's the reason effort to rid the world of superstition and religious myths or it's a method for testing about the physical world through observation analysis and experimentation or it's all the above I won't have a quiz but definition c is is the one that i think is the best s best definition of science definition a is is something that often happens we do get natural explanations about the world around us through the process of testing and observation and experimentation but it's not limited to just that if you were an environmental chemist and you're trying to to look at the distribution of lead in the ocean you would be making decisions about whether the concentrations that you saw were totally natural whether they were pollution which would be anthropogenic man derived or some combination so you're actually discerning the difference between natural and other processes definition b is sort of a red herring i just threw that in the sea of people were strongly biased against superstition and religious myths of course if either A or B or C is not there that it can't be D but method C really is all about how science is done it is a method for testing our ideas and we compare them against the physical world we want something that is unbiased that doesn't have philosophical overturns we want something that that we can use as a as a yardstick yeah uses a yardstick for comparison science is also not about believing you know this science is about observation and experimentation then we really know what happens when we have a certain set of conditions you're not extrapolating you're not now imagining what happens between things you know belief on the other hand requires that we have faith in things unseen or trust and things hope for that's an entirely different set of standards it doesn't mean that belief is wrong and that science is always but there are two different things and clearly you know science works to this different standard where we test our ideas versus reality so why am i as a chemist interested in or speaking to you tonight about the origin of life now well if abiogenesis is about the origin of life you know shouldn't we be listening to a biologist well before life begins there's only chemistry and maybe some physics but there's no biology so you really do need a chemist to look at this issue chemistry is also a repeatable thing chemicals don't evolve if you have a set of chemicals now they're the same as they were a million years ago and they'll be the same as it will be in nine years in the future they'll do the same thing all the time this makes chemistry testable so we can really test the ideas we have about how life originated some of the ideas that people are considering and testing is that life is the result of unguided chemical reactions and random processes another idea is life is the result of purposeful physical laws and initial conditions in other words somebody stack the deck and set things up for life to happen a third idea is that the laws of nature and the fine-tuning of the initial conditions are insufficient to account for the origin of life something has to happen continually along the way they just can't stack the deck the way you actually have to cheat as you go along or limiting our options to natural processes alone prohibits the origin of life these are all ideas we can test through experimentation one of the most famous experiments about the origin of life is the miller-urey experiment in 1953 Stanley Miller and he really stunned the world with the results of his experiment where he announced that amino acids could be made from a mixture of reducing gases with an electric spark this this coldly caught the world by surprise and subsequent to his initial analysis people have identified in that mixture all the sugars all the nucleic bases a lot of the fatty acids the things that you need is basic building blocks for assembling a cell virtually everything you want to find was in that mixture now the news stories describing this work you know they appeared on the front page of newspapers worldwide now big banner headlines life in a test-tube this really caught the attention of the world and the experiment in most cases was hailed as the first step in the origin of life by a purely naturalistic pathway people finally thought that they had found the key to the origin of life this is sort of a schematic of the process that they were envisioning in the first step here this is what Miller did into his experiment he took a simple mixture of gases and produced this complex mixture of fatty acids and amino acids sugars and cheering's and pyrimidines then the idea was that these compounds would pulverize the fatty acids would would react with glycerin and make triglycerides than these would be the basis for lipids amino acids were polymerized the peptides and eventually a change would be long enough to be called proteins and enzymes the sugars they would combine to make complex carbohydrates things like starches and cellulose and of course the periods and pyrimidines these are the building blocks of RNA and DNA and then it was envisioned that by some process that they didn't quite know all these things would get together and eventually make a true cell and at first glance this seems very reasonable it's it's a very simple system it's very elegant instant simplicity you might even think that so simple and so straightforward that it was designed which is probably the worst thing you can say about a natural process now at the time George Wald wrote an article for Scientific American where he picked up on the Miller experiment and sort of elaborated it as a possible pathway and the first step in the pathway to the origin of life and had some quotes from his his article and what he said was however improbable we regard this event or any of the steps which involved it involves given enough time that will almost certainly happen at least once you know he was totally convinced that once you make the basic building blocks things would all come together he says that time is in fact the hero of the plot the time which we have to deal is on the order of two billion years later in the article he said what we regard is impossible on the basis of human experience is meaningless here in other words forget everything you know given so much time the impossible becomes the possible the possible probable and the probable virtually certain one only has to wait time itself performs the miracles which is a very curious thing for a scientist to say as for he's probably just being very flamboyant here but he's talking about miracles and that gives you an idea of how complex this process really is because it's something that we wouldn't normally see we wouldn't normally envision happening he said time was enough once we had those basic building blocks we only had to wait well not everybody was convinced by his article a few years later about a decade there's an anonymous editorial that appeared in nature and they took a totally different view of the situation they said that those who work on the origin of life must necessarily make bricks without very much straw which goes a long way to explain why this field of study is so often regarded with deep suspicion speculation is bound to be rife and has also frequently been wild and what they're saying here is there's not very much data that's the bricks without straw so there's a lot of speculation and people are speculating quite wildly further than the article they said that some attempts to account for the origin of life on Earth however ingenious I've shared much with imaginative literature and little with theoretical inference of the kind which can be confronted with observational evidence of some kind another in other words it's all science fiction it's not been tested in the laboratory so in fact when you go back and you look at this this scheme for abiogenesis this is the part that Stanley Miller did but nobody at the time a while writing his article knew how any of these steps of have happened and today it's still pretty much that case there are a lot of question marks you don't know how these things are happening in fact the more you look at this figure the more it looks like this cartoon we're have a lot of stuff at the beginning you have what you want at the end but in the middle you have a miracle happening about 25 years after he wrote that article Scientific American did a compendium of different articles on the origin of life and when they reproduced or reprinted Wald's article there was an editorial that preceded it that basically retracted what he said and what they said in the editorial is that although stimulating this article probably represents one of the very few times in his professional life when Wald has been wrong examine his main thesis here and see can we really form a biological cell by waiting for chance combinations of organic compounds sir Fred Hoyle lichen such an occurrence - the assemblage of a 747 by a tornado whirling through a junkyard from what I've seen a tornado's this is not likely to happen later in the editorial they said Harold Morra what's in his book energy flow in biology computed that merely to create a bacterium this would be one of the simplest organisms will require more time than the universe might ever see if chance combinations of its molecules were the only driving force that's not they're not talking about the time that has preceded us but he's talking about the whole life time in the universe would not be sufficient time for chance combinations and that's a little hard to think about so what I thought I would illustrate is how many chance combinations of amino acids there are now in Miller's experiment he found about seven or eight amino acids but by the end of the 70s about 50 different amino acids had been found and all but two have at least two optical isomers and some of them have several centers of optical isomerism so you have 2 4 and 8 possibilities for those amino acids and you need a minimum chain length of about a hundred residues before a protein begins to have enzymatic activity and for each amino acid there are at least two ways of reacting they have an amino group they have a carboxylic acid group they can form the peptide bond but they could also turn around and form an acid anhydride they could also react with side chains for the ones that have amino or acidic side chains and the longer the chain gets the more possibilities you have but what I did for this calculation we just say there's just two ways we'll keep it simple so the number of possible combinations if you do the math you've got the 50 amino acids the two optical isomers the 2 ways of combining and you need a hundred of them in a chain so you work through the math and you can lop to a number that's to two hundred and thirtieth power if you're not familiar with scientific notation that's a very big number to give you an idea how big that number is what if we just had one of each of these molecules how much would that whole thing weigh well to get the weight we take the number of molecules times the average molecular weight of the protein and divided by Avogadro's number so the average molecular weight of an amino acid is about 130 140 that's 100 times the average weight and then when you put these amino acids together you make 99 peptide bonds so we have to take out 99 times 18 which is the weight of water that comes out from that peptide bond and when you go through and do the math the the average weight of one of these 100 amino acid peptide proteins is a hundred eight thousand 218 Dalton's to get how much they all weigh together we take the 10 to the 230th power times the weight divided by Avogadro's number and we get an answer that's ten to two hundred and tenth power grams that's a that's a lot to give you an idea how much a lot it is the mass of the universe the whole universe not just the known universe but with everything including the dark matter is on the order of 10 to the 56 grams so we would need in excess of 10 to the hundred-and-fifty power universes that mass to make all these amino acids so we had just one of all the possible combinations so random chance combinations of amino acids produces a huge number of possibilities but only some of them are going to have enzymatic activity and and those are the ones that you want you have to come out of this big mass that's going to be a difficult problem but what really killed Walt's thesis wasn't calculations like this but it was a discovery of new evidence and it was startling new evidence between the early 50s and the the middle delayed 70s bark horn and shop had discovered in rocks micro fossils fossils of what looked like bacteria and algae and these were in rocks three and a half billion years old more recently deposits with carbon isotopes characteristic of living organisms were found in formations 3.8 billion years old the problem you have is that instead of having four and a half billion years to 3.8 to make life the earth wasn't habitable for the first seven hundred million years depending on how fast it formed there there are different ideas but in something and most of these you have the earth its surface being too hot now you don't even have liquid water it's so hot then you have another problem that around the four billion the 3.8 billion year time period there was what's known as the late heavy bombardment looking at cratering on the moon there was this the second episode after the planets and the moon and everything had formed there was the second episode of a very large meteor impacts and it's thought that they may have been sufficient to melt the surface of the earth or if not at least to make it uninhabitable by life so you you don't begin to look for life to form on the earth until about 3.8 billion years ago but we have evidence that life may have been existent almost instantaneously if and if that if the carbon isotope work turns out to be invalid we have visible algae and in other micro fossils in rocks at 3.5 billion years so most you have is 300 million years you don't have the two billion years that we all talked about and I have no doubt that is as more discoveries are made this this time window for life to originate is going to get smaller so there's this exceptional little time for life to form by unguided chemical reactions and that's why Wald's idea the time was sufficient to perform the miracles was was dismissed currently when people talk about the origin of life what they're trying to do is to give us a what they call a coherent narrative they know that they're having problems identifying what the early Earth was like but they want to give us this this the story if you will of how life originated now that's nice but if we don't check that story against reality it's just a story and we're trying to do a scientific evaluation here and stories just aren't going to be good enough we need to be able to test these things now if you want an idea where things stand presently there's a very nice book written by Jeff bethe and Christopher Willis and it's called the spark of life Darwin and the primeval soup and in there they've collected a lot of the current thought and a lot of the current work on the origin of life but it's presented as a nice coherent narrative they don't do a scholarly criticism of this work this is a popular book they don't want people to get all wrapped up in controversy they just want to lay out how it might have happened on the other hand there are people that like the the nature editorial aren't buying it a friend of mine and a member of my thesis committee Ken Nealson he said nobody understands the origin of life if they say to do they're probably trying to fool you when I found this quote from Ken I thought well that's that sounds rather cynical and maybe he didn't say it cuz I found it on the internet so I wrote to Ken my ass and did you really say this and he said he did and well I probably not as cynical as kin is about this that they're probably trying to fool fool us they may have managed to fool themselves and that's something we all have to be careful about especially in in an area like this where there's so little that we can actually test our ideas against so the question is how do we get into this mess where we've got some scientists that say it's not a problem it all works out there are all these possible reactions and in the end they're all the compounds come together to form a cell and on the other side of the fence there are scientists saying well nobody knows what they're talking about they're all just trying to fool you how we got in this mess is actually a long story but I'll try to be brief the concept of spontaneous generation is a very old one the Greeks consider this a possibility empedocles was first proposed that life arose naturally he envisioned this as a continuous ongoing process it doesn't happen just once it happens continually and that the forms have persisted were those best adapted for survival and reproduction which sounds very familiar to me but we won't worry about that tonight his ideas were repeated and expanded upon by epicurus and lucretius other Greek philosophers but Plato and Aristotle rejected this idea that life is constantly forming they saw design and purpose in nature and for them design implied a designer now jumping ahead a couple thousand years we notice a Darwin when he wrote his book Origin of Species did not include any discussion of the origin of life and that seems to me like a rather big hole given the title of his book did he simply duck the question or maybe you just thought well it's outside the scope of my discussion or did he consider the origin life a given since at the time the concept of spontaneous generation was still a viable idea people still thought that life arose spontaneously what we know is that the Origin of Species was written in 1836 to 1844 wasn't published until 1859 but the experiments that Pasteur did to disprove spontaneous generation were not done until 1862 1864 so it's possible Darwin consider that the spontaneous origin of life was something very possible what we do know is that in 1871 seven years after Pasteur published his results disproving spontaneous generation Darwin wrote a letter to his friend Joseph Hooker where he said it is often said that all the conditions for the first production of a living organism are now present which could ever have been present but if I know what a big if we could conceive of some warm little pond with all sorts of ammonia and phosphoric salts light heat electricity present that a protein compound was chemically formed ready to undergo still more complex changes at the present day such matter would be instantly absorbed which would not have been the case before living creatures were formed so he's a vision you get this warm little pond with just the right ingredients for making a cell and they all just come together now it was another 50 years before anybody produced anything of substance along this line and what happened is 18 sorry 1924 or pyrin and in 1928 Haldane apparent is a Russian scientist and all vain is a British biologist separately and independently proposed the under reducing conditions on the earlier early Earth amino acids might be formed and somehow this prime mortal prebiotic soup life somehow emerges so they envision a reducing atmosphere giving rise to Darwin's warm little pond well it wasn't for another 25 years before somebody actually tried to test the idea to actually see is will this really work and that's the now-famous miller-urey experiment where they did indeed make amino acids using reducing conditions now what they were trying to do in this experiment will simulate the prebiotic chemistry on the early earth and by prebiotic chemistry we just mean the chemistry that happens before life begins they took apart and Haldane's idea of a reducing atmosphere and they applied an energy source to drive the organic compound synthesis and they built this apparatus to simulate the early Earth and their idea was you put the ingredients in you turn on the electric spark wait a few days and in the end it would produce the building blocks of life and they really expected to see just about every possible organic molecule come out of this now what are reducing conditions and if we imagine that the foremost well you don't have to imagine we know the four most important elements to life are hydrogen carbon nitrogen and oxygen in a reducing atmosphere the hydrogen would be in the form of molecular hydrogen it would be a gas carbon would be in the form of methane nitrogen will be in the form of ammonia and oxygen would be in the form of water vapor our current oxidizing atmosphere hydrogen is in the form mostly of water there's there's some trace levels of hydrogen but these are being continually produced and they're rapidly consumed carbon is in the form of co2 nitrogen is the molecular nitrogen gas and oxygen is molecular oxygen gas so what they were what Miller and Urey did is they use this reducing mixture and this is the apparatus they designed it has a large flask a five liter flask where they had their atmosphere they were the two tungsten electrodes to make the spark that would simulate lightning they had a condenser to simulate the cloud forming processes and rain they had a little trap here that would be filled with fresh water condensing from the from the large bulb this would be like the fresh water streams running down the sides of the continents and then they had an ocean here where they would be drawing their samples out where the organic compounds should collect if they were formed and they heated this flask so that the water vapor would go back up and circulate and this would go around and round and when they did the experiment they really did find all the building blocks of life they found amino acids they found fatty acids not real long-chain ones they found things like acetic in and pro prion ik they found many of the nucleobases that you need for RNA and DNA and they found some small sugars but if you read that miller's papers very carefully you'll also notice that he mentions this oily red goo coating the walls of his apparatus it turns out this is the major carbon product of the experiment but most people don't concern themselves with it because it's not one of the building blocks these are what they were interested they focused on and they just sort of ignored this for for years we'll come back to this later now as I said earlier the experiment was hailed as the key step leading to the natural origin of life it was frequently imitated there was actually a small cottage industry of maybe 25 or 30 scientists right after Miller did his experiment trying to reproduce it and they used a variety of different reducing atmospheres a variety of energy sources and the products that got were largely the same there were some changes in distribution but they all got amino acids they all got fatty acids they all got some of the purines and pyrimidines it seemed like this was a general process that happens whenever you have a reducing atmosphere and they were all convinced that the first hurdle to forming life abiotic li was had been crossed but the question is did it really happen that way now is there any evidence on the earth that that's how life actually originated or that we even had this time of prebiotic synthesis and it turns out there's no rocks or sediments from this time of about four billion years ago so there's no way we can test whether it actually happen but there's also some rather serious problems with the miller-urey experiment the amino acids fountains were in very low abundance given that analytical techniques of the time Stanley Miller was lucky to find them he used paper chromatography his detection limits are thousands of times higher than what you can determine today with a gas chromatograph the most most of the amino acids that he found have turned out to be non protein amino acids these are amino acids that organisms do not use they have different structures and all the amino acids that he produced were racemic in living organisms today only the elf form is used and the reason for this is proteins the the peptide chain is a is a coil and if you have a mixture of the dnl form you can't form that coil it kinks it turns all different ways what Miller did not find was any peptides he didn't get any homogeneous polymerization of the amino acids and this homogeneous polymerization of amino acids to form peptides its Thurman DiNapoli unfavored even when you have pure substances in solution it's even worse when you mix the amino acids with other compounds and most recently the presence of a reducing atmosphere when the early Earth has been questioned some dismiss it as completely unlikely such as Howard Holland at Harvard and the problem we have there is we just don't have any real evidence of what the atmosphere was like on the early Earth but the people haven't given up last year in science Channel had a new model for the atmosphere on the early Earth where they were able to produce a hydrogen-rich atmosphere so this is a question that is still actively being debated debated today and they'll fight forever because they don't have any samples to solve the question we also don't know what the conditions on the early Earth were like was it freezing was was the accretion here so slow that it was cold was it fast and we had a molten surface of the earth or was it something in between there just isn't evidence to resolve these questions now shortly before I became a graduate student at Scripps the Murchison meteorite it fell in Australia this was the year man landed on the moon and although this meteorite landed way in the Australian outback the people there knowing that the United States was sending men to the moon they thought this this meteorite would be very interesting to them so when they collected the meteorite they they kept it clean they kept it uncontaminated and they shipped it off to NASA Ames here in California and also to the Field Museum in Chicago they found several different pieces of it this meteorite business was identified as a carbonaceous chondrite these are these meteorites the carbonaceous chondrites are the oldest material in the solar system and it's thought that they may contain some evidence that will help us to decide about life's origin and shortly after it was sent to NASA Ames the team that was supposed to look at the moon rocks for organic compounds got this me to write and they analyzed it it's a case of the prepared now chance favoring the prepared mind they were ready to look at really low levels and a sample just walked in their door one day and what they found were very low abundances of amino acids we're talking a few micrograms per gram of meteorite so a couple of parts per million and what they found in this pattern of found in the amino acids was a pattern very solar similar to the miller-urey experiment mostly the amino acids were non protein amino acids which is also very similar to the miller-urey experiment and all the amino acids were receiving so if you had an optical Center you've got both the D and L forms but the most abundant form of carbon in the Murchison meteorite was a dark red to black tar most people don't pay much attention to that because it's not exciting like amino acids so at the time that this discovery was announced Stanley Miller and Jeff bethe they came up with an idea and they said well the striker cyanohydrin pathway was postulated as the way amino acids were formed in the miller-urey experiment but they noticed that this pathway is forked the noble path when there's lots of ammonia present you get only amino acids but there's an alternate path and if there's no ammonia present the pathway produces hydroxy acids so what they suggested is is if the ammonia concentration on the immediate parent body was some moderate amount of ammonia then both products the high amino acids and the hydroxy acids would be formed and they could use the ratio of the appropriate pairs to tell us about the ammonia centration on the meteorite parent body all they needed was two things mom was the sample of the Murchison meteorite and the other was a graduate student that they could get to do the work and the ivory members distinctly the day Jeff proposed this in class and he baited the hook and I bit hard it seemed like a really exciting project to work on and being a first year graduate student needing a thesis project it was all just laid out there for me to take and so I went I talked to him and he agreed to to let me work on the project what I didn't know is going to take me three years to develop the analytical method the hydroxy acids are a lot more difficult to work with in amino acids they're actually quite volatile and so it was a painstaking process of developing the method but I've actually got it to work and I was all excited I went to Jeff and I said well I'm ready to go let's get the let's get the sample the meteorite and do the analysis and he said well we better call Stanley Miller first and talk to him and Miller Wiley's wisely suggested that I test the method when in one of the experiments that he had run some of that extract from the flask and see if I could find hydroxy acids in there it should be about the same degree of difficulty and then they could really test to see if my method did work wasn't that they didn't trust me but there are many Murchison meteorites in the world and you don't want to blow your one good sample the the method worked very well in the miller-urey experiment I got to do the meteorite and the assay for hydroxy acids in the Richards amiright worked very well we found an abundance pattern identical to what I found in the miller-urey experiment the hydroxy acids were all racemic so I knew they were non-biological origin they weren't contamination from my fingerprints and things like that and we were able to use the appropriate pairs of amino and hydroxy acids to calculate the ammonia concentrate and on the meteorite parent body and it came out the one or two millimolar which doesn't mean a lot to you folks but it was very close to a number the jet calculated on his thesis research about what the concentration of ammonia should be on the earth due to the stability constants of some of the amino acids so we're all very pleased and happy this was this was great exciting work but it didn't generate anywhere near the fanfare that either the middle of your experimented or the discovery of amino acids in a meteorite nobody knows what hydroxy acids are and they really don't care so I wisely decided that well the reason I was at Scripps was to get a PhD in work in oceanography so I I would pursue my career as an oceanographer and not worry about analyzing meteorites for the rest of my life that turns out to be a good decision because carbonaceous chondrites don't fall that often about once every 30 years it's hard to build a career when you have 30 years between samples so everything seemed to be well at the when I finished my degree but over the years I begin to think about some of the things and some of the implications of this work now the Miller bethe prediction that we would find hydroxy acids in the immunity that was confirmed and this striker sign a hydrant pathway has been well identified as a source of the amino acids in the meteorite and also as a possible path white one the early Earth or is it the question of the reducing atmosphere is is really key to this if you don't have a reducing atmosphere this pathway doesn't happen now at the time that that this reducing atmosphere was became disputed nor said well not to worry we've got meteorites now that make these and we find these amino acids and hydroxy acids and meteorites maybe the meteorites bring them to the earth and we don't have to have a reducing atmosphere so he did the calculation based on estimates of the flux of meteorites and the concentration of amino acids in the merchant cemetery and he was very disappointed because it turns out that if you take all the amino acids and all the meteorites and put them in the Earth's ocean you only get about ten billions of amino acids ten millionths of a gram per liter this is a very small concentration nothing is going to happen at those little concentrations what people said not to worry the world's oceans are actually that water was brought here by comments and comments are going to be loaded with organic compounds and we just have to analyze the comments and and everything will be fine well we're still waiting for the common analysis so we haven't tested that idea yet but if we look back at the miller-urey experiment there's another problem now why are the results from the Murchison meteorite and the Miller experiments so similar as the meteorite it's four and a half billion years old it represents untold years of chemical synthesis maybe millions maybe billions we don't know the experiment that Stanley Miller did was only six or seven days now why should the two look so very familiar and the other question that that lingered the one that I've struggled with is what is the source of this red goo and electric discharge experiment and the black tar in the Murchison meteorite now these are the major forms of carbon in both cases where are they coming from you know when in economics people say if you want to figure out what's going on you follow the money or in politics for that matter well in chemistry if you want to figure out what's going on particularly with origin of life you've got to follow the carbon so we need to look at where are the major products the this red goo and the black tar where are they coming from so I've looked at Miller some of his old data from one of his early papers and this this shows the kinetics of the reaction and you can see as time goes by ammonia concentration it drops pretty linearly and shortly after the experiment starts hydrogen cyanide and aldehydes are produced but they quickly plateau they reach the steady-state concentration and short while after the amino acids start to come in but they plateau as well and these these plateaus here these are what chemists call steady states and what's happening is this experiment really involves three steps the first step is the synthesis of the aldehydes and hydrogen cyanide from the gases these aldehydes and hydrogen sining then react through the striker cyanohydrin pathway to make amino acids hydroxy acids the third step is that something is consuming the amino acids because if you've got one process producing them you have to another one consuming them at balanced rates to get that steady-state concentration but there's no evidence of polymerization of the amino acids to peptides we don't find peptides in this experiment so the question is are the amino acids reacting with other compounds and forming something else and the answer is yes it's it's called the Maillard reaction and it's actually familiar to all of us here's peppermint pattie and she's working on her science project and she had some trouble but she actually came up with something she made toast now it's it's that's a rather simple science project but she was very proud of it and it's not the baking or the making of toast but rather it's the baking of the bread for the toast that the Maillard reaction is very important it was first described by lewis camille meillard in 1912 and what happens is the amino acids from protein they interact with reducing sugars like glucose or lactose and this is why you add milk when you bake bread and not table sugar which is not a reducing sugar and these these compounds reacts to produce colors aromas and flavors characteristic of a particularly cooked food now temperature accelerates them the Maillard reaction products give bread crusts cakes and cheese and pizza sorry cheese pizzas their distinctive brown color and the small volatile compounds that are produced as a byproduct of this reaction are responsible for the the camel like in most of the Roma's that you get when you bake bread and baked pizza now for the chemists I threw in this slide the rest of you don't panic it's not as bad as it looks but here's the reducing sugar reacting with an amino acid and it's the amino group that attacks the reducing part of the sugar for you know through the measured reaction producing a Midori compounds and these then undergo a whole cascade of different reactions giving different products one of these is a sticker striker degradation where you lose co2 amines and you get back to aldehydes that you started with when you were making amino acids all of these products then combined to produce melanoidins now mil anoints is this large class of compounds that have one thing in common that's they're all colored that's why they're called milla noids it's sort of a one of those trashcan terms as you can throw a lot of things into you don't know what's in there but they're all colored I looked up the yeah Maillard reaction on the internet a few years ago to see how widely it was studied and it turns out it's a big deal this was the seventh international symposium it was held in Tokyo and and a few other cities in Japan what was interesting is the two groups of people that were coming you can't read the sessions here - well but there are two kinds one is medical science and the other is food science I would imagine this be a great conference to go to because all the food scientists they're going to they're going to be interested in good food so the meals they are going to be fabulous and if you're over in Joel indulge there's all these medical doctors there that'll take care of you as I said earlier temperature accelerates the Mayo reaction that's why we we heat food to cook it but it can also proceed at room temperature and I think you've all seen this happen as well if you slice an apple and just let it sit in a few hours it turns brown and what's happening there is that when you cut the Apple you destroyed cells and release a lot of compounds they're now free in that apple juice that you're creating to react on their own they're no longer bound up and they produce these colored compounds during that same time frame if you had a mixture of amino acids you're not going to make any peptides and if you cook something too long you get melanoidins and black tars won't burn toast this reaction is also very important in aging now proteins and carbohydrates the lens crystalline protein that turns brown as people get old this is part of cataract formation is you're forming these brown colored compounds via the Mayo reaction in your eyes and products of the mirror reaction have been isolated from Alzheimer's disease lesions they've also been implicated in arterial sclerosis and diabetes mellitus in other words we are slowly cooking as we age and as a consequence of disease I know it's not a pleasant thought but it's one of the things that happens to us the real question is why don't we cook faster the Apple after we sliced it it only took an hour to turn brown our body is in a much higher temperature why isn't this reaction just stopping our biology cold and the answer is it's due to the cell structure amino acids and sugars inside a cell they're not floating free there are all kinds of internal organizations within a cell that contain the various substances we have ribosomes to control protein synthesis and this is mitochondria to facilitate the energy reactions and it's these structures that are keeping the sugars and the amino acids apart so what we're seeing is the complex structure of the cell itself is needed to make the components necessary for making the cell and keeping them from reacting uncontrollably or randomly so how does this all relate back to buy a biogenesis well we don't know how any of these reactions could possibly be occurred on the early Earth but we do know that if you've got a mixture that's composed of this it's going to react and produce mil annoyed and eventually with enough cross-linking going to go to karagin this is the the red oil that miller saw is the experiment and this is the black tar that we find in the meteorite so we know from what observations we made that the second step in the miller-urey experiment was not the production of biopolymers but were these colored what we call geopolymers this means that the mälar reaction is really a showstopper in terms of abiogenesis and it's not just in the in his experiment and in the meteorite that this reaction is important if one studies organic geochemistry you'll probably see this picture somewhere in your class and what happens when organisms die their cells break apart they release the car bhai some lipids all these biopolymers are released into the environment through microbial degradation and also in organic decomposition these bio polymers are broken down into their component parts sugars organic acids amino acids fatty acids this mixture should be very familiar to us by now this is what Miller produced in the natural environment today these compounds are reacting through the Mayo reaction producing complex assemblages of problem arising you've got fulvic acids being produced you make acids and as they continue to crosslink they produce the geopolymers the melon oeid karagin that we find in rocks and sediments so the difference between biopolymers and geopolymers you know structure-wise biopolymers are linear proteins are a linear chain of amino acids in the geopolymers things are branched they go every which way and we have all kinds of Bond types here we have just the peptide bond the components well you know in proteins we have amino acids in DNA and RNA we have the nucleic acids in starches and cellulose we just have sugars so it's just a single compound class in the geopolymers they're all mixed up together functionally the biopolymers act as catalysis they were sort of a form of energy storage and their starches and they make up our genome the geopolymers it stars that they have no no chemical functions in terms of information content there's all kinds of stuff stored in our genome in the DNA and RNA there's no information in the geopolymers so what does all this apply for the miller-urey experiment well the amino acids that we found are intermediates in the reaction pathway not the end products and what we find are actually remnants of the reaction and that's why you find the low yield the prebiotic synthesis makes melanoidins this is the early red goo that you saw in the flask from the building blocks of life prebiotic chemistry is essentially over as soon as it starts you don't make the biopolymers that you need no matter how much time you have because you're making the melanoidins compare this to what you find in a in a single-cell organism single-cell organisms like prochlorococcus which is a marine photosynthetic prokaryotic an make all the chemicals it needs to make an exact copy of itself this single cell is you know it's capable of all the complex biochemistry that's essential for life and cellular structure is essential to the process without this cellular structure as we saw with the Apple when you slice it the Maillard reaction takes over and makes melanoidins so with the structure it's possible to make the homogeneous polymers that we need the cell function grows and reproduces itself we're now at a point where we can say that well the cell is an irreducibly complex structure you can't make it piecemeal only cells can make cells this was something that that Pasteur was proving in his early experiments life was coming from life it wasn't forming spontaneously so we've got this cell that the only way we can make it is the processes in that cell making a copy of itself so a quick summary so far despite the apparent success of the miller-urey experiment in 1953 many problems remain re-examination of this experiment shows that this pathway is not the road to Darwin's warm little pond nor is it a pathway to biomolecules or microbes yeah think of what happened with that Apple we're going to be making melanoidins not microbes on the other hand complex cellular structures are essential there are an essential characteristic of and a prerequisite to life you've got to have these things so the origin of life requires a level of complexity the undirected chemistry cannot achieve regardless of how much time is available we've got this chicken or egg situation so we need the cell to make the compounds that are in it and we need the compounds or in it to make the cell which came first biologists recognize how difficult this problem now is if M Harold in an article in 1995 says the origin of life stands as the most profound mystery in biology and we note in passing that despite all the achievements of investigators in prebiotic biochemistry it remains utterly beyond our comprehension that's a fancy way of saying what Ken Nealson said earlier nobody knows how to do this lynn margulis who was one of the main proponents of the abiogenesis she admits that to go from a bacterium to people is less of a step than to go from a mixture of amino acids to a bacterium just recognizing that this first step the first living organism is the most difficult problem because of these obvious problems I think if we're honest we will recognize that abiogenesis is it best it's a myth of modern science it's a nice coherent story that we tell ourselves but at worse it's a lie we tell ourselves so we can pretend we know more about the origin of life than we actually do we are actually fooling ourselves when we say that this is the process that life began so what's the chemist to do about this well they've tried lowering the bar let's not try and make the whole cell let's try and make just different parts well synthesized in the building blocks that's done but now we've got to separate the building blocks to keep them from reacting chemists can do this acting intentionally but how does this happen in nature what kind of random process pulls up pulls a mixture apart another simple step would be to make some basic bio polymers but how does that happen in nature when if you put the polymers themselves in water they hydrolyze and come apart then the idea is well once we've got the basic thing started some sort of chemical evolution will occur and will evolve more complex molecules now I have to say honestly now that I'm not a graduate student thinking about this it's probably the most crazy thing I ever heard molecules don't evolve they're chemicals they react and then become another chemical but to evolve means that they have some sort of reproduction they don't and the last step was a symbol of minimal cell we can't make a complex cell let's try and make something very basic so these are all ideas that people are trying to use today to advance the idea of abiogenesis one of the big ideas that's being pushed is these self-replicating molecules another idea is that proteins came first the idea is once you have some proteins maybe they helped make the DNA maybe proteins and nucleotides appeared together and they work and helped each other out maybe a metabolism came first maybe membranes came first and there are other ideas let's just look at this these first five these are ideas of breaking the process down into smaller steps Leslie Orgel is one of the big promoters of self-replicating molecules and this is an idea that he has proposed now he hasn't done the experiment this is just an idea that needs to be tested okay you have this molecule down here that because of its various functional groups attracts these other two molecules together and when these two groups react and these two groups should be very familiar because here's the amino group and the aldehyde and this combination here is the first step in the maelard reaction they joined together and now you have a molecule that's identical to the one you started with now the curious thing here is this looks like a great idea but remember these are the two functional groups that make the mälar reaction so this group can react with any aldehyde so if it's not bound up here and it's floating out there's other competitive reactions that are going to be going after it as well and the same is true for this aldehyde function or go also very cleverly use the same names for the side chains R 1 R 1 R 2 and R 2 so that his condensed molecule will be identical to the starting one but there's absolutely no way that this molecule controls what these side chains are these could be anything so it's not really a self-replicating molecule it is that actually just a template that might help other molecules come together what he's done here is a semantic illusion I don't know whether he's intentionally trying to trick us or whether he's managed to trick himself as well but this isn't a self-replicating molecule the idea the proteins first and proteins and nucleotides together mind fret agonist is one of the the big proponents of this as I his thoughts are the ends attack enzymatic activity is first can aid the synthesis of the genetic material well we have these problems there's no evidence that proteins form naturally and in the experiments that he's done to get these things to happen he's done we're like what has been called investigator intervention this is things like eliminating all the competing reactions so you only put amino acids in when you're trying to do a polymerization it's things like adding leaving groups to the ends of the molecules so they join together and the reaction actually happens and it's other things like putting blocking groups on the molecules where you don't want them to react so they join together the right way and not the random way which is what would be happening in nature if he wasn't there he's worked with Freeman Dyson one is this double origin of both the clack acids and proteins and the idea is there's the synergy between the two compound classes but if we think about it without the protection of cell wall both materials will be consumed by them a large reaction so these are nice ideas but when you really put them in nature there are the competing reactions that are very fast that are going to al compete to help catalyze these process Karen Smith is said well maybe clays are somehow involved in these organic compounds were absorbed on the surface of clays and and that gets the thing going well just a week or two ago David Deamer there was an article in science about David Deamer from UCSC he added a mixture of proteins DNA and cell membranes to a hot little puddle in Kamchatka Russia and when I first looked at this picture is a little hard to see here he is pouring a beaker of some liquid I thought well is he just pouring a quart of milk in but reading the news article with it they actually had this mixture of the he made up of proteins DNA and cell membranes well where did they come from I mean shouldn't you be starting with amino acids nucleotides and fatty acids I mean he's he's a couple steps ahead already well what happened is an experiment was that the compounds he added within a few hours were totally absorbed by the Clay's and that's where they stay they were completely taken out of solution and they were done nothing happened besides the Clay's absorbing the organic compounds so clays don't actually help catalyze these processes what the Clay's do is they strip things from solution and this the prebiotic soup just gets more diluted with time Harel Maura which is a big proponent of the reverse tricarboxylic acid cycle this is the cycle that cells used to generate energy and he uses this to explain the production of long-chain fatty acids that can be used to make cell walls well this process involves the directed input of hydrogen and carbon dioxide and there's no way to contain the product so even if it works a couple times around as you start to build up a longer longer chain it's going to drift away you need a cell to keep the products there also many of the products of this process are subject to degradation by the Maillard reaction you know that that reaction is just killing everything because these are their fatty acids they have the ketones and aldehydes functions that can be attacked by amines the other process is is naked undirected count or you know make it undirected chemistry chemistry outside of cell it's not cyclic chemicals react and they go downhill thermodynamically and they go downhill very fast but they don't go around in cycles another idea is that well maybe have membranes first and you get things called these protocells or coacervate says they used to be calm and they capture chemistry and take it inside well that's a nice idea but as soon as the chemistry inside the cell reacts it's done you'd have to worry about the fusion through these cell membranes to get more supplies in to keep the thing going if the cell was or this membrane was was porous enough for things to come in well all kinds of things would come in and you'd pretty soon have the mayor reaction going on inside the cell so what is happening in all these examples is people are just grasping at straws many of these problems that they had before still remain bio polymers are not formed by random reactions the various compound classes must be separated or they're going to react through the mayor of reaction and consume each other the biopolymers have to be assembled in very specific ways you know proteins require a peptide bond they're just not any old random linkage the bio polymers are composed of hundreds of mala Murs monomers arranged in a specific order there's information content in these molecules like I showed earlier if if you just do it randomly there's a vast number of combinations but only a few of them are ones that actually work and cells require hundreds to a few thousands of enzymes in specific places the genetic material must be isolated and the energy compounds must be utilized properly this is a complicated structure it's just not a bag of loose chemicals Bruce Alberts now current in' and that well former head of the national academy of science and a well known biologist he says that the cell is a biochemical assembly line you know it's a little factory all the factories I know have been designed so what we need to do is we need to recognize that what prebiotic chemistry produces the melanoidins and the geopolymers is really of no use to the origin of life and what we need the biopolymers of specific compositions prebiotic chemistry cannot produce well results so far indicate that life is not the result of unguided chemistry processes it just hasn't happened life is also not the result of purposeful physicals and initial conditions you know we've put these chemicals together and they're just not doing what we need them to do now the laws of nature and fine-tuning of the initial conditions we need to recognize that they are insufficient to account for the origin of life limiting our options to just natural processes alone in fact inhibits the in hard origin of life you're just not going to get there with what natural random reactions do well if nature is insufficient then maybe it's time to consider other options one of the ideas as crazy as it sounds that's that's actually becoming very popular today is that life came here from outer space it was first proposed by Svante Arrhenius and he proposed the bacteria being pushed by light beams came from other planets to the earth this idea was recently championed by Fred Hoyle and Wickramasinghe and also by Francis Crick now the problem with it is that's how life came to earth is it may explain why there's life on the earth but it doesn't tell us where life came from because it puts the origin of life out there somewhere on another planet where we can't even begin to look at the conditions so it's a I find it a very unsatisfactory explanation and there's all kinds of problems with getting things here through space which I don't have time to go into but if we're so desperate that scientists are seriously considering panspermia maybe it's time for us to consider other options as well maybe it's time to consider design now if the cell is a miniature biochemical assembly line then maybe it was designed by a prebiotic intelligence directed chemistry can make all the biopolymers but we don't know which to make a priori you know as a graduate student I was able to separate all the compound classes to do the analysis today graduate students they can make enzymes full working functional enzymes people are trying to assemble viruses from scratch so we're at the point where we can make the bio polymers that are necessary the trick is people are just copying what's already been done and we're not randomly putting these things together we're putting them together matching patterns that we already find in nature so we're not smart enough to know how to put them together and once we've got the compounds we're not smart enough how to know to put them back into a cell you know when people do tissue experiments and they extract them or put them in a tissue miser and they break up all the cells and they want to pull out the different components nobody ever worries that they're going to spontaneously go back together this isn't something that just happens the cells have to be carefully assembled and we just don't know how to do that so if cells are done by an intelligence it's an intelligent smarter than we are and clearly that's what's needed I'll conclude with with two thoughts one is from Karl Popper now he said this over 30 years ago it's even more true today and that's the unjammed of breakthroughs in molecular biology has made the problem of the origin life a greater riddle than it was before we have acquire new and deeper problems you know what Miller was trying to do 50 years ago has now gotten harder the complexity of cell is way beyond what we knew then the other advice that will leave you with was written by saul of tarsus in the first century test everything hold on to the good now i've thrown out a lot of ideas here tonight I encourage you to or challenge you to test them hold onto the good ones throw out the bad ones but don't just take my word for it there's some investigating for yourself thank you very much mic while they're pondering I'll put up a suggested reading list for people that want to investigate some more these are five books that they might find of interest you mentioned it a number of times but you didn't really go into it the genetic code I mean of all the things that you point out it seems the most wondrous is that we can have the level of information that we have in the genetic code put together in a certain sequence not only of its own right but the fact that it can also then template all protein and all other manufacturing that takes place in a cell in the DNA that I mean that the amount of information content in there is larger than maybe in all the books and all the libraries in the world in one particular cell in that one string of DNA ten to the ten bases and I wonder if you'd like to comment about that well there's the the information content is is really overwhelming that it's in it's in a really single molecule the other thing about the genetic code that that is very curious is there is no chemical reason for why the base pairs code for different amino acids this is a code that could have been the written anyway that the code does code for specific amino acids now but it doesn't have to be the way it is because because of the way it's transcribed the RNA molecules they pick a particular amino acid but that could be assembled anyway this is this is a code that is purely arbitrary in development and those kind of codes speak to me of intelligence now there is no underlying physical reason why a certain triplet of base pairs codes for a particular amino acid this this is something that was assigned early on and whenever I see a code well I think of a code writer yeah one thing I was very interested in is if you had ever come across any attempts to explain the fact that all of the amino acids and proteins are left-handed instead of right-handed by by naturalistic processes okay that's a that's a very good question it's one that there are no satisfactory answers for you'll see it in lots of places where people saying we still don't know why it's just L amino acids in the book by Willis and beta they talked about a scientist that did an experiment where he built an enzyme using all d amino acids and found that it worked on the mirror image substrate that the L protein did so there there is no I priority chemical reason why it's all L or all D it has to be but it has to be one or the other because if you want to build a coil of amino acids they all have to go in the same direction likewise with DNA and RNA you use D sugars and the reason for that is with the D sugars the 5 carbon is above the plane of the sugar and the 3 hydroxy group is below so you can build your phosphate bridges this way and have the base pairs come off if you use L sugars it goes the other way so you can go in the other direction but if you mix them up you can't build that nice chain I'm nucleotides and makes us the helix thanks I have one more question I'm going to I got the impression from the summary at the end of your talk that you don't hold out much hope I mean if there's avenues most of the common avenues that could possibly have been in place on the early earth in the in the pre cellular earth have already been explored and they haven't led anywhere I mean so is this field going anywhere are there people that are still actively pursuing things or are we at sort of the point of view of just postulating well we don't know how it happened it just happened and we're going to leave it there state I mean where are where is this field well is to feel going anywhere are there people still active up to questions yes it's still active there are still people trying to do these kind of experiments I'm not impressed by the results because frequently if you look at them carefully there's a lot of investigator intervention you know they're stacking the deck for things to happen and they're doing things that are not conceivable on the early Earth you don't find amino acids with leaving groups and blocking groups so that they'll form proteins the way you want them to so yes there's people still active but I don't think it's going anywhere I think it's it's run its course I think they've missed a lot by not paying attention to the fact that in the miller-urey experiment the walls were coated with polymers that were forming as fast as the amino acid were being produced they've missed an essential part of the chemistry of the early Earth if you're going to have a synthesis in a reducing atmosphere you're going to produce all these compounds in a mixture and it's going to produce this red oily goo and that's going to be its major product and what it does is it takes the vast majority of the compounds that you produce and it makes the red oil eventually it becomes a tar that's not going to leave much left to build things with it's been 50 years since the miller-urey experiment it's still the premier experiment and I think it's time for people to look at this in the harsh light of day and say we're not getting anywhere I think back to the problem with alchemy where 500 years BC it was proposed that there were four elements earth air fire and water and these were everything was made from these four substances and so you could take lead and make gold if you just took out the right amount of of earth and put in enough fire or whatever 2,000 years went by a failed experiments trying to do this and finally somebody said well you know maybe letting gold are elements themselves and we can't make one from the other and it was that idea that quickly led to atomic theory and the modern periodic table that never would have happened if they just stuck with the four elements and I think what's happening with a bio chemistry resource of prebiotic chemistry research is they're still in that alchemical stage where they're looking for the magic combinations of the four gases and or some other equivalent of that and they're not thinking outside the box that more is new than just natural reactions they need information and they need to direct that synthesis and those aren't natural processes but those are that's what's needed to get a cell does does a specimen this chemistry have anything to do with the specified complexity in the DNA know that the complexity of DNA you know that's that's a code that's written in you could by changing the the RNA that takes that reads the code and is transferred to where the proteins are made because they carry an amino acid with them you could take the part that reads the code and the part of the molecule that carries the amino acid and you could put them together any way you wanted to you could change that code be a lot of work but but there's no reason why a certain triplet codes for a certain amino acid other than the fact that it does and because of that it codes for the RNA that carries that transfer you know it's it's information that's built into the code but it's not required by any chemistry so chemistry couldn't really produce the information no and and like the amino acids the the number of combinations of nucleotides that you could come up with is enormous it seems you alluded to the problem and it may be well understood I'm just not that familiar with it but one of the critical things is that you also need an RNA to transcribe the DNA if you ever got something like your 100 molecule chain is there any accepted model for why you for how the RNA would have developed to transcribe that information and actually give you a replicating system if you even got the necessary complexity in the first molecule the simple answers is no one of the things that Orgel is promoting with the RNA world is that RNA is both the memory compound and the transcriber you know that way he's lowered the bar with the first organism is just a pure RNA organism and it has RNA doing the memory work as RNA doing the transcribing work and he may even have some RNA doing some enzymatic work this seems to him a simpler task to achieve but it creates a whole new set of problems because how do we get from that RNA world to our current combined DNA RNA world that that's that's that's just as big a problem as creating one or the other and didn't it wasn't that the slide in which you pointed out that he had no control over the side chains right but there's another question that interests me that even if you assume that somehow that came to exist is there any if you think of the problem of assembling such a molecule the first time is there any kind of estimate of the time that it would take to form such molecules or do those are those of reasonable simplicity were that they could fit into that sort of 300 million year window the only estimate I know of the time involved was Harold morrow it's one where he said it the whole time of the universe is greater than what we need to to get these things happening by random reaction it's it's a big problem and mostly people say well we got lucky we got really really lucky if it happened by random chance it makes the lottery look like a sure thing intelligent design may be something that the philosophers of science are able to develop a coherent research program around but do you think that that intelligent design is are you proposing that become part of science or something that simply gives perspectives to the science that we do in other words it might be a useful religious or a worldview or philosophical idea that makes sense of the world the science we do describes the world that we live in but it cannot answer certain questions how how do you think intelligent design and design and science work together order or don't they well I think it's still early to say exactly how it's all going to come together I think there are some research programs that intelligent design will will push forward there will always be the charge that it's religiously based but limiting ourselves just to natural reactions or natural processes and saying that nature is all there is is philosophically based currently science has been abducted if you will by naturalist and they've closed the doors to thinking outside their box it's causing some problems specifically looking at this at this question of the origin of life it's pretty clear natural reactions aren't capable of doing what we need to be done and what they do is not what we want and so we need to get outside that box one way to do that is to say that there is more or there was more operating at the origin of life than just the natural relaxed reactions and we can see how design processes can can accomplish many of the tasks what you may not be able to say is well did this really happen this way we have to go we have we have the problem we can't go back to the to the past and look at it and we don't have much evidence but it does present a process that is capable of accomplishing the task the other issue with with intelligent design is it doesn't specify the designer but many of the opponents of intelligent design insists that it's being presented as a way of sneaking the the conservative Christian God in the back door of science class let's turn that around if if we work insisting that intelligent design pointed to the conservative Christian God we would never be able to defend that position because there is so little about the designers that revealed in this process that it'd be hard to make that conclusion justify it but it's one that's been hung on the ID movement by the other side I think the resolution of the problem is to say there is evidence or how does one decide about evidence for design we need to look at that question and then ask okay now that we know how to identify design do we find evidence for it in nature and biology and that's a that's a two-step process it's we're just at the beginning of the process of identifying design and that's mostly a mathematical process at this point but as that develops it will then allow asking the second question and those are valid questions that can be asked and studied scientifically by experimentation I think that's where it may help us well thank you all for coming and thanks to the Veritas forum for inviting me it's been a real pleasure for me and thanks for sticking out I know it went a little long you you

Channel: Access Research Network

Views: 15,645

Rating: 3.8571429 out of 5

Keywords: Abiogenesis, origin of life, Edward Peltzer, Naturalism (Idea), Creation--evolution Controversy (Legal Subject), Stanley Miller

Id: 6xj4UH0RwcM

Length: 94min 38sec (5678 seconds)

Published: Sun Feb 09 2014