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Davisson, J. A.

Publications and source records attributed to Davisson, J. A..

2 recordsLinked to original sources

Nonenzymatic, prebiotic aminoacylation couples chirality of RNA and protein

Life as we know it depends on the homochirality of nucleic acids and proteins. However, there is no widely accepted explanation for why life uses only D-sugars for nucleic acids and L-amino acids for proteins. Here we demonstrate a prebiotically plausible method of nonenzymatic aminoacylation in a water ice-eutectic phase. These reactions produce high yields of aminoacyl-tRNAs, which are active in translation. Surprisingly, we discovered these nonenzymatic aminoacylation conditions were stereoselective, favoring coupling of amino acids and RNA of "opposite" L- and D- configurations. D-RNA shows greater aminoacylation yields for L-amino acids. The opposite was true for L-RNA, which had greater yields with D-amino acids. Nucleic acid backbone chirality influencing stereoselectivity of aminoacylation presents the missing link in the origin of modern biochemistry. This phenomenon provides insight into the chirality of the RNA world, and helps to explain the "opposite" stereochemistry of modern biomolecules.

biochemistry↗

S1 hydrophobic residues alter the voltage sensing phosphatase enzymatic function and voltage sensing

The voltage sensing domain (VSD) is a four-helix modular protein domain that converts electrical signals into conformational changes, leading to open pores and active enzymes. In most voltage sensing proteins, the VSDs do not interact with one another and the S1-S3 helices are considered mainly as scaffolding. The two exceptions are the voltage sensing phosphatase (VSP) and the proton channel (Hv). VSP is a voltage-regulated enzyme and Hvs are channels that only have VSDs. To investigate the S1 contribution to VSP function, we individually mutated four hydrophobic amino acids in S1 to alanine (F127, I131, I134 and L137). We also combined these mutations to generate quadruple mutation designated S1-Q. Most of these mutations shifted the voltage dependence of activity to higher voltages though interestingly, not all substrate reactions were the same. The kinetics of enzymatic activity were also altered with some mutations significantly slowing down dephosphorylation. The voltage dependence of VSD motions were consistently shifted to lower voltages and indicated a second voltage dependent motion. Co-immunoprecipitation demonstrated that none of the mutations broke the VSP dimer indicating that the S1 impact could stem from intrasubunit and/or intersubunit interactions. Lastly, when the same alanine mutations were introduced into a genetically encoded voltage indicator, they dramatically altered the optical readings, making some of the kinetics faster and shifting the voltage dependence. These results indicate that the S1 helix in VSP plays a critical role in tuning the enzymes conformational response to membrane potential transients and influencing the function of the VSD.

biophysics↗