Evolutionary optimization of a nonbiological ATP binding protein for improved folding stability

Chemistry & Biology
John C Chaput, J W Szostak

Abstract

Structural comparison of in vitro evolved proteins with biological proteins will help determine the extent to which biological proteins sample the structural diversity available in protein sequence space. We have previously isolated a family of nonbiological ATP binding proteins from an unconstrained random sequence library. One of these proteins was further optimized for high-affinity binding to ATP, but biophysical characterization proved impossible due to poor solubility. To determine if such nonbiological proteins can be optimized for improved folding stability, we performed multiple rounds of mRNA-display selection under increasingly denaturing conditions. Starting from a pool of protein variants, we evolved a population of proteins capable of binding ATP in 3 M guanidine hydrochloride. One protein was chosen for further characterization. Circular dichroism, tryptophan fluorescence, and (1)H-(15)N correlation NMR studies show that this protein has a unique folded structure.

References

Feb 9, 1978·Nature·W Gilbert
Jan 1, 1972·Methods in Enzymology·Y Nozaki
Mar 11, 1994·Science·R D JenisonB Polisky
Jan 1, 1995·Methods in Enzymology·R W Woody
Oct 1, 1995·Nature Structural Biology·A R DavidsonR T Sauer
Jan 1, 1995·Annual Review of Biochemistry·R F Doolittle
Aug 5, 1993·Nature·M Sassanfar, J W Szostak
Dec 15, 1994·Nature·C A OrengoJ M Thornton
Mar 15, 1994·Proceedings of the National Academy of Sciences of the United States of America·A R Davidson, R T Sauer
Oct 1, 1995·Protein Science : a Publication of the Protein Society·J R Desjarlais, T M Handel
Jan 1, 1997·Annual Review of Biophysics and Biomolecular Structure·C ChothiaA Murzin
Sep 1, 1997·Nucleic Acids Research·S F AltschulD J Lipman
Aug 15, 1997·Structure·C A OrengoJ M Thornton
Oct 6, 1997·Science·B I Dahiyat, S L Mayo
Nov 14, 1997·Proceedings of the National Academy of Sciences of the United States of America·R W Roberts, J W Szostak
Oct 1, 1995·Chemistry & Biology·B E EatonD A Zichi
Jan 8, 1999·Journal of Molecular Biology·C Zhang, C DeLisi
Sep 29, 1999·Proceedings of the National Academy of Sciences of the United States of America·M W WestM H Hecht
Nov 11, 1999·Journal of Molecular Biology·S JungA Plückthun
Mar 15, 2000·Journal of Molecular Biology·G ChoJ W Szostak
Jun 29, 2000·Annual Review of Biochemistry·W F DeGradoA Lombardi
Jul 13, 2000·Methods in Enzymology·R LiuR W Roberts
Mar 29, 2001·Proceedings of the National Academy of Sciences of the United States of America·D S WilsonJ W Szostak
Mar 29, 2001·Proceedings of the National Academy of Sciences of the United States of America·C A VoigtZ G Wang
Apr 5, 2001·Nature·A D Keefe, J W Szostak
May 25, 2001·Nature Structural Biology·D VitkupC Sander
Nov 17, 2001·Chemical Reviews·D A Moffet, M H Hecht
Jun 25, 2002·Chemistry & Biology·Michael McPhersonBrent L Kreider
Mar 26, 2003·FEBS Letters·Tomoaki Matsuura, Andreas Plückthun
Apr 3, 2003·Chemistry & Biology·Shuwei Li, Richard W Roberts
Nov 1, 2003·Proceedings of the National Academy of Sciences of the United States of America·Yinan WeiMichael H Hecht
Nov 25, 2003·Science·Brian KuhlmanDavid Baker

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Citations

Dec 22, 2006·PloS One·Yuuki HayashiTetsuya Yomo
Jul 11, 2008·Expert Opinion on Biological Therapy·John C ChaputBerea A R Williams
Jul 17, 2012·Trends in Biotechnology·Agathe UrvoasPhilippe Minard
Jun 23, 2007·Journal of Molecular Biology·Sheref S MansyJohn C Chaput
Feb 18, 2010·Protein Science : a Publication of the Protein Society·Junko TanakaHiroshi Yanagawa
Jul 30, 2005·Journal of Molecular Biology·Michael WunderlichFranz X Schmid
Feb 16, 2005·Expert Opinion on Biological Therapy·Maria A T Groves, Jane K Osbourn
Nov 14, 2014·PloS One·Aleardo MorelliBurckhard Seelig
May 9, 2008·Chembiochem : a European Journal of Chemical Biology·Jennifer L Watkins, John C Chaput
Sep 17, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Li Zhang, John C Chaput
Jun 2, 2005·Protein Engineering, Design & Selection : PEDS·Nobuhide DoiHiroshi Yanagawa
Jun 16, 2009·ACS Chemical Biology·Chad R SimmonsJohn C Chaput
May 7, 2021·Theory in Biosciences = Theorie in Den Biowissenschaften·Athel Cornish-Bowden, María Luz Cárdenas
Jan 1, 2020·ACS Synthetic Biology·Matilda S NewtonBurckhard Seelig
Jun 15, 2006·Journal of the American Chemical Society·James M CarothersJack W Szostak

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