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Antibody catalysis of peptidyl-prolyl cis-trans isomerization in the folding of RNase T1

Ma, Lifu and Hsieh-Wilson, Linda C. and Schultz, Peter G. (1998) Antibody catalysis of peptidyl-prolyl cis-trans isomerization in the folding of RNase T1. Proceedings of the National Academy of Sciences of the United States of America, 95 (13). pp. 7251-7256. ISSN 0027-8424. PMCID PMC22581.

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An antibody generated to an alpha-keto amide containing hapten 1 catalyzes the cis-trans isomerization of peptidyl-prolyl amide bonds in peptides and in the protein RNase T1. The antibody-catalyzed peptide isomerization reaction showed saturation kinetics for the cis-substrate, Suc-Ala-Ala-Pro-Phe-pNA with a k(cat)/K-m value of 883 s(-1) M-1; the reaction was inhibited by the hapten analog 13 (K-i = 3.0 +/- 0.4 mu M). Refolding of denatured RNase T1 to its native conformation also was catalyzed by the antibody, with the antibody-catalyzed folding reaction inhibitable both by the hapten 1 and hapten analog 13. These results demonstrate that antibodies can catalyze conformational changes in protein structure, a transformation involved in many cellular processes.

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Additional Information:Copyright © 1998 by the National Academy of Sciences. Contributed by Peter G. Schultz, April 22, 1998. We thank Elizabeth Sweet and Yoko Oei for assistance with antibody production. This work was supported by the National Institutes of Health, predoctoral fellowships from the National Science Foundation and American Chemical Society (L.C.H.-W.), and a postdoctoral fellowship from the Cancer Research Fund of the Damon-Runyon Walter Winchell Foundation (L.M.). P.G.S. is a Howard Hughes Medical Institute Investigator and a W. M. Keck Foundation Investigator. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:NMR-spectroscopy, isomerase, protein, cyclophilin, ribonuclease-T1, amide, complexes, collagen, FK506, assay
PubMed Central ID:PMC22581
Record Number:CaltechAUTHORS:MALpnas98
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:1006
Deposited By: Tony Diaz
Deposited On:29 Nov 2005
Last Modified:19 May 2015 00:06

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