Adam and Eve? Or Alanine and Serine?

What if the defining reason for life on Earth is the manipulation of time? While organic matter has been discovered in outer space, in the form of methane in the ethers of Neptune and Uranus, and asteroids and meteorites that crashed into Earth (Organic Compounds in the Solar System, 2016), why have we not yet discovered extraterrestrial life in the form of a cell that is moving, eating, growing, changing, and replicating (Sagan & Sagan, 2018)?

The defining feature of life could be the enzyme: proteins made from amino acids that increase the rate of chemical reactions. Metabolic reactions could not occur without enzymes (Lewis & Stone, 2020), as reactions would occur too slowly to interact with one another. Enzymes increase the speed of reactions so that complex, multifaceted interactions can occur to create and sustain life. It is the enzyme that has catalyzed life on Earth, by shortening the time needed for reactions to take place.

Although amino acids have been discovered in some meteorites, such as the simpler alanine and glycine found in class CI and CN meteorites (Organic Compounds in the Solar System, 2016), the 20 main ones that define life on Earth have not yet been found elsewhere in the universe in same concentrations and varieties.

There is much debate surrounding the origin of the 20 amino acids. Is it by chance or by evolutionary design? And are more amino acids evolving or could they evolve? Other amino acids do exist, including selenocysteine which is found in humans, but is comparatively more complex to utilize, which might be a clue as to why there aren’t more than the 20 main ones that exist (Brazil, 2017).

Francis Crick hypothesized in his frozen accident theory that 20 specific amino acids exist by chance and that any other number of structures could have been used to synthesize proteins (Koonin, 2017). However, in 2017 Doig makes strong arguments that the 20 main amino acids with their specific hydrophobicity enable folding, stability, accessibility of active sites, and metabolic efficiency, making amino acids the perfect building blocks for proteins and enzymes (Doig, 2017).

Why are C, H, N, O, and S the organic molecules that compose amino acids? In addition to being bountiful on our planet, these elements are ideal for many reasons. In contrast, metals such as selenium and antimony are too soluble in water and therefore unstable. Halogens are too electronegative and reactive. Silicon bonds too readily to oxygen in place of other elements. The list goes on and on to account for the specificity that explains why our current amino acids are the perfect constituents of life (Doig, 2017).

So, if it is not a frozen accident then why has the number of amino acids stopped at approximately 20 when so much of evolution is characterized by seemingly endless diversity? One fascinating theory explains it in terms of simple logistics. tRNA (transfer RNA) reads the genetic code from RNA, and then using the enzyme aminoacyl tRNA synthetase, selects which amino acid tRNA should bind to. tRNA then transfers the selected amino acid to a ribosome for assembly on the growing polypeptide chain. tRNA has only 3 reading sites, and after accounting for the start and stop codons, 61 possible amino acids reading codes. However, tRNAs are limited in their recognition ability (Saint-Léger et al., 2016). tRNAs already make on average one mistake for every 1000-10,000 codon readings and adding more amino acids would enable more mistakes. Dr. Ribas explains it this way, “It’s like if you have a very simple kind of lock where you could only change three or four pins, you come to a point where you wouldn’t be able to make new keys because a new key will open a lock you have already used and that defeats the purpose” (Brazil, 2017).

Photo Credit: (National Human Genome Research Institute, 2019)

Scientists do not have all the answers on whether amino acids evolved by chance or limited necessity, but one thing is certain; the 20 amino acids are brilliant in their specificity and enable life as we know it. Without proteins, we would not have enzymes, and without enzymes, we would still be a pool of RNA and co-factors. Enzymes alter the speed of reactions and amino acids are the specific building blocks of all proteins and enzymes. Without our 20 amino acids, we would lack the complex web of reactions needed to create life; organisms that can move, eat, breathe and reproduce.

 

REFERENCES

Organic Compounds in The Solar System. (2016, May 26). Chemistry LibreTexts. https://chem.libretexts.org/Ancillary_Materials/Exemplars_and_Case_Studies/Exemplars/Physics_and_Astronomy/Organic_Compounds_in_The_Solar_SystemLinks to an external site.

Sagan, D., & Sagan, C. (2018). life | Definition. In Encyclopædia Britannica. Retrieved from https://www.britannica.com/science/lifeLinks to an external site.

Lewis, T., & Stone, W. L. (2020). Biochemistry, Proteins Enzymes. PubMed; StatPearls Publishing. https://www.ncbi.nlm.nih.gov/books/NBK554481/#:~:text=Enzymes%20are%20proteins%20comprised%20ofLinks to an external site.

Brazil, R. (2017). Why are there 20 amino acids? Chemistry World. https://www.chemistryworld.com/features/why-are-there-20-amino-acids/3009378.articleLinks to an external site.

Doig, A. J. (2017). Frozen, but no accident – why the 20 standard amino acids were selected. The FEBS Journal, 284(9), 1296–1305. https://doi.org/10.1111/febs.13982 Links to an external site. 

Koonin, E. (2017). Frozen Accident Pushing 50: Stereochemistry, Expansion, and Chance in the Evolution of the Genetic Code. Life, 7(2), 22. https://doi.org/10.3390/life7020022 Links to an external site. 

Saint-Léger, A., Bello, C., Dans, P. D., Torres, A. G., Novoa, E. M., Camacho, N., Orozco, M., Kondrashov, F. A., & Ribas de Pouplana, L. (2016). Saturation of recognition elements blocks evolution of new tRNA identities. Science Advances, 2(4). https://doi.org/10.1126/sciadv.1501860Links to an external site.

National Human Genome Research Institute. (2019). Transfer RNA (tRNA). Genome.gov. https://www.genome.gov/genetics-glossary/Transfer-RNA

The past and future of aminoacyl-tRNA synthetases

I chose aminoacyl-tRNA synthetases (aaRSs) to study today because these enzymes seemed like some of the more complex and mysterious molecules from this week’s lectures on protein synthesis. The more I researched, the more I understood how one could completely dedicate one’s life to studying only one enzyme. I read an in-depth paper by Rubio Gomez and Ibba (CSHL Press) and was surprised to discover that there are currently 23 known aaRSs. In addition to the 20 that Goodsell references that each code for a specific amino acid, there are two that code for lysine and two called pyrrolysyl-tRNA synthetase and phosphoseryl-tRNA synthetase found in some archaea and bacteria (Goodsell) (Rubio Gomez and Ibba).

Interestingly, aaRSs presents some interesting questions regarding their evolution, and, as others have mentioned, there are rare genetic disorders linked to aaRs mutations, as well as recent aaRs-targeted drug developments. Interestingly, these enzymes play a central role in the latest biosynthetic research as engineered amino acids are being written into novel polypeptide chains, altering the genetic code and resulting in the creation of biosensors, biomarkers, innovative functioning proteins, viral defenses, and more. Scientists have altered some aaRs:tRNA pairing to associate a new amino acid into translation (Rubio Gomez and Ibba) (Rovner et al.). As there is so much to discuss regarding aminoacyl-tRNA synthetases, I will focus on how they evolved and how they are altering evolution.

AaRSs are antiquated, having been inherited from the Last Universal Common Ancestor (LUCA) (Fournier et al.). And, they seem to have been just as complex in LUCA as they are in contemporary organisms (Rubio Gomez and Ibba). Since aaRSs are the readers of the genetic code, but also the genetic code is required to synthesize them, this presents a ‘chicken or the egg’ dilemma (Rubio Gomez and Ibba). As discussed in lecture 39, there are two types of aaRSs, each approaching the tRNA from a different side with their active site, with type I adding an amino acid to the last 2’ tRNA hydroxyl group and type II adding an amino acid to the 3’ hydroxyl on the final tRNA base (Barbaro). Both types also have a difference in their substrate binding methodology. Despite their difference, there is an accepted theory called the Rodin and Ohno hypothesis that both classes arose from the same gene simultaneously from opposing sides. The codons that code for residues for class I active sites are palindromes for class II sites. In other words, codons for class I are anticodons for class II. (Martinez-Rodriguez et al.). And so, two aaRSs arose due to bidirectional reading of mRNA and attached two different amino acids to tRNAs. This created the first protein comprised of more than one amino acid. Later genetic mutation and editing enabled the diversity of present-day aaRSs (Rubio Gomez and Ibba).

Noncanonical amino acids (ncAAs) are artificially synthesized amino acids that alter the genetic code and create genomically recoded organisms (GROs). GROs are created when scientists reassign a codon to a different and sometimes artificially synthesized amino acid (Lajoie et al.). Thus, new aaRSs are required that can recognize ncAAs and attach them to tRNAs that will transport them to ribosomes for protein chain synthesis. For example, Rovner et al. conducted research on an organism that lacked the TAG codon and reassigned TAG to code for a Methanocaldococcus jannaschii tRNA:aminoacyl-tRNA synthetase pairing. The protein was fabricated and stable.

I was curious to understand how aaRSs are altered to bind nCAAs. I came upon a study that described exactly its methodology and provided a helpful infographic. If you take a look at part c 2. (top right), you can see that a library of aaRS variants is generated after the introduction of the novel amino acid. Through mutation, an aaRS will mutate to associate the codon with the ncAA. This will be selected and amplified using PCR technology (Vargas-Rodriguez et al.). I was surprised that ncAAs are created by selective breeding! 

Image: www.ncbi.nlm.nih.gov/pmc/articles/PMC6214156.

I have only scratched the surface of my understanding of aaRSs. From ancient origins to the future of genetic engineering, it seems there is ad infinitum to discover in the study of aminoacyl-tRNA synthetases. 

 

 

REFERENCES

Rubio Gomez, Miguel Angel, and Michael Ibba. “Aminoacyl-tRNA synthetases.” RNA (New York, N.Y.) vol. 26,8 (2020): 910-936. doi:10.1261/rna.071720.119

Fournier, Gregory P., et al. "Molecular Evolution of Aminoacyl tRNA Synthetase Proteins in the Early History of Life." Origins of Life and Evolution of Biospheres, vol. 41, no. 6, 2011, pp. 621-632.

Barbaro B, Biochemistry at U of California San Diego. Course Number: Chapter 39 - the Genetic Code” [accessed 2023 Dec 15]

Martinez-Rodriguez, Luis, et al. “Functional Class I and II Amino Acid-activating Enzymes Can Be Coded by Opposite Strands of the Same Gene.” PubMed Central (PMC), 18 June 2015, www.ncbi.nlm.nih.gov/pmc/articles/PMC4528134 Links to an external site..

Rovner, Alexis J., et al. “Recoded Organisms Engineered to Depend on Synthetic Amino Acids.” PubMed Central (PMC), 21 Jan. 2015, www.ncbi.nlm.nih.gov/pmc/articles/PMC4590768 Links to an external site..

Goodsell, David. “PDB101: Molecule of the Month: Aminoacyl-tRNA Synthetases.” RCSB: PDB-101, pdb101.rcsb.org/motm/16. Accessed 16 Dec. 2022.

“Genetically Modified Organisms (GMOs) | Learn Science at Scitable.” Genetically Modified Organisms (GMOs) | Learn Science at Scitable, www.nature.com/scitable/topicpage/genetically-modified-organisms-gmos-transgenic-crops-and-732. Accessed 16 Dec. 2022.

Lajoie, Marc J., et al. “Genomically Recoded Organisms Expand Biological Functions.” PubMed Central (PMC), www.ncbi.nlm.nih.gov/pmc/articles/PMC4924538. Accessed 16 Dec. 2022.

Vargas-Rodriguez, Oscar, et al. “Upgrading aminoacyl-tRNA Synthetases for Genetic Code Expansion.” PubMed Central (PMC), 27 July 2018, www.ncbi.nlm.nih.gov/pmc/articles/PMC6214156.

 

Where does the fat go when you lose weight? Out your mouth? Really??

This is my response to a biochemistry assignment, in which our professor asked us to read and respond to the following article: https://www.medicalnewstoday.com/articles/287046

This is wild because just two days ago I bought a Lumen, which is a device that you breathe into throughout the day that claims to track your respiration exchange rate (RER), which is your CO2 to O2, exhalation/inhalation ratio. Lumen claims their device can train your metabolism by providing personalized advice on exercise and diet in response to your RER. This week’s assignment is a perfect opportunity to dig a little deeper into the relationship between RER and weight loss and see if this was a smart purchase.

Image from https://www.lumen.me/metabolic-flexibilityLinks to an external site.

The specific statement I would like to focus on from the article “Majority of weight loss occurs ‘via breathing’” is:

The results suggest that the lungs are the main excretory organ for weight loss, with the H20 produced by oxidation departing the body in urine, feces, breath and other bodily fluids.

This statement is derived from the works of Brown and Meerman, as published in their peer-reviewed article in the Britsh Medical Journal. The authors deduce that the equation for the oxidation of a single triglyceride is:

C55H104O6+78O2→55CO2+52H2O+energy

Triglyceride breakdown equation from Meerman and Brown, 2014.

Using stoichiometry, Brown and Meerman explain that 10 kg of fat requires 29 kg of oxygen which results in 11kg of H20 and 28 kg of CO2. Building on the 1949 research of Lifson et. al., which found that the synthesis of carbonic acid powers the exchange between oxygen in H20 and gaseous CO2, Meerman and Brown traced oxygen’s path and found that for every 4 O2's exhaled, 2 bind with hydrogens to form water. Therefore, according to Brown and Meerman, exhalation is indeed the main exit pathway for oxygens broken up by triglycerides, with the remainder exiting as H20, in a 2:1 ratio (Meerman & Brown, 2014).

These findings are reinforced by another peer-reviewed article that is essentially a lab directive for students to calculate oxygen and food intake against carbon dioxide output after exercise (Merritt, 2022). I found an interesting peer-reviewed paper on bats that found that the exhalation ratio in bats reflects the food type (protein, carbohydrates, fat) their muscles are currently burning (Youngsteadt, 2011). There are many articles that measure metabolic adaptation by way of carbon dioxide exhalation monitoring, such as the 2018 study by Moll et al., which found CO2 exhalation to be a reliable way to observe aerobic and anaerobic pathways in athletes, in addition to blood lactate concentration testing.

After my research, I am convinced that carbon dioxide exhalation is the exit pathway for catabolized triglycerides. However, I would like to see more articles that directly test this hypothesis.

Many questions remain. Can the Lumen breath analyzer show reliable readings that accurately reflect what the metabolism is burning? While metabolic flexibility (the ability to shift between fat burning to carbohydrate burning, for example) is affirmed by research (Goodpaster & Sparks, 2017), the Lumen itself is not proven to be a trustworthy measurement device (Hall, 2021).

Luckily, the Lumen has a money-back guarantee. I will test it out for myself and let you know my anecdotal experience!

 

REFERENCES

Meerman, R., & Brown, A. J. (2014, January 1). When somebody loses weight, where does the fat go? The BMJ. Retrieved November 22, 2022, from https://www.bmj.com/content/349/bmj.g7257Links to an external site.

Lifson, Nathan, et al. "The fate of utilized molecular oxygen and the source of the oxygen of respiratory carbon dioxide, studied with the aid of heavy oxygen." J Biol Chem 180.2 (1949): 803-11. https://web.archive.org/web/20171023163139id_/http://www.jbc.org/content/180/2/803.full.pdfLinks to an external site.

Merritt, Edward K. "Why is it so Hard to Lose Fat? because it has to Get Out through Your Nose! an Exercise Physiology Laboratory on Oxygen Consumption, Metabolism, and Weight Loss." Advances in Physiology Education, vol. 45, no. 3, 2021, pp. 599-606. https://go.exlibris.link/4sb7LsvF Links to an external site. 

Youngsteadt, Elsa. "Bats Gorge during Exercise."American Scientist, vol. 99, no. 2, 2011, pp. 124. https://go.exlibris.link/W60561rX Links to an external site. 

Moll, Kevin, et al. "Comparison of Metabolic Adaptations between Endurance‐ and sprint‐trained Athletes After an Exhaustive Exercise in Two Different Calf Muscles using a multi‐slice 31P‐MR Spectroscopic Sequence."NMR in Biomedicine, vol. 31, no. 4, 2018, pp. n/a. https://pubmed.ncbi.nlm.nih.gov/29393546/ Links to an external site. 

Goodpaster, B. H., & Sparks, L. M. (2017, May 2). Metabolic Flexibility in Health and Disease. Cell Metabolism. Retrieved November 23, 2022, from https://www.cell.com/cell-metabolism/abstract/S1550-4131(17)30220-6Links to an external site.

Hall, H. (2021, July 27). Lumen’s Information Is Not So Illuminating. Science-Based Medicine. Retrieved November 23, 2022, from https://sciencebasedmedicine.org/lumens-information-is-not-so-illuminating/ Links to an external site.