Engineering cyclophilin into a proline-specific endopeptidase

Nature
E QuéméneurA Ménez

Abstract

Designing an enzyme requires, among a number of parameters, the appropriate positioning of catalytic machinery within a substrate-binding cleft. Using the structures of cyclophilin-peptide complexes, we have engineered a new catalytic activity into an Escherichia coli cyclophilin by mutating three amino acids, close to the peptide binding cleft, to form a catalytic triad similar to that found in serine proteases. In conjunction with cyclophilin's specificity for proline-bearing peptides, this creates a unique endopeptidase, cyproase 1, which cleaves peptides on the amino-side of proline residues. When acting on an Ala-Pro dipeptide, cyproase 1 has an efficiency (kcat/Km) of 0.7 x 10(4) M(-1) s(-1) and enhances the rate of reaction (kcat/kuncat) 8 x 10(8)-fold. This activity depends upon a deprotonated histidine and is inhibited by nucleophile-specific reagents, as occurs in natural serine proteases. Cyproase 1 can hydrolyse a protein substrate with a proline-specific endoprotease activity.

References

Mar 1, 1992·European Journal of Biochemistry·W Bode, R Huber
Jun 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·J Liu, C T Walsh
Dec 14, 1990·Science·D L ScottP B Sigler
Apr 7, 1988·Nature·P Carter, J A Wells
Nov 1, 1980·Analytical Biochemistry·T H Plummer, M T Kimmel
Jan 1, 1995·Progress in Biophysics and Molecular Biology·A Galat, S M Metcalfe
Apr 1, 1995·Toxicon : Official Journal of the International Society on Toxinology·B MaillèreA Ménez
Jan 6, 1995·Science·A Radzicka, R Wolfenden
Jan 19, 1995·Nature·H J DugglebyP C Moody
May 10, 1994·Proceedings of the National Academy of Sciences of the United States of America·B W MatthewsH Neurath
Nov 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·E A Merritt, M E Murphy

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Citations

Jul 9, 1999·Journal of Molecular Recognition : JMR·A HuberD Neri
Aug 10, 2000·Journal of Molecular Recognition : JMR·A Skerra
Oct 7, 1998·Protein Science : a Publication of the Protein Society·E Farinas, L Regan
Apr 2, 1999·European Journal of Biochemistry·H Czapinska, J Otlewski
Nov 4, 2011·International Journal of Phytoremediation·Joseph M Jez
Nov 7, 1998·Clinical Chemistry and Laboratory Medicine : CCLM·J B Weitzman, M Yaniv
Nov 29, 2001·Proceedings of the National Academy of Sciences of the United States of America·D N Bolon, S L Mayo
Jun 7, 2000·Proceedings of the National Academy of Sciences of the United States of America·D E BensonH W Hellinga
Mar 31, 1999·Proceedings of the National Academy of Sciences of the United States of America·A E NixonS J Benkovic
Jan 6, 2001·Memórias do Instituto Oswaldo Cruz·A R TeixeiraJ M Santana
Sep 2, 1999·Protein Engineering·P Iengar, C Ramakrishnan
Apr 24, 2007·Angewandte Chemie·Miguel D ToscanoDonald Hilvert
Nov 27, 1998·Nature Biotechnology·M C FletcherJ S Lee
Mar 27, 2007·Protein Science : a Publication of the Protein Society·Michael HulkoJörg Martin
Mar 21, 2003·Protein Engineering·Mariana Peimbert, Lorenzo Segovia
Oct 3, 1998·The Journal of Biological Chemistry·W R Cannon, S J Benkovic
Jul 18, 2006·The Journal of Biological Chemistry·Ben Youngblood, Norbert O Reich
Feb 24, 2017·Journal of Biosciences·Michel Morange
Apr 20, 2001·Progress in Biophysics and Molecular Biology·N LahavA C Elitzur

Dec 29, 2020·Biologie aujourd'hui·Pierre AntonyChristophe Romier
Jan 9, 2008·The Journal of Organic Chemistry·Xiyun ZhangK N Houk
Apr 1, 1999·Chemistry & Biology·P J O'Brien, D Herschlag
Nov 17, 2001·Chemical Reviews·T M Penning, J M Jez
Nov 12, 2002·Journal of Combinatorial Chemistry·Annemieke MadderPierre J De Clercq
May 26, 2007·Biochemical and Biophysical Research Communications·Shuang LiJin Zhang

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