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Originally published In Press as doi:10.1074/jbc.M104362200 on September 27, 2001

J. Biol. Chem., Vol. 276, Issue 48, 44481-44487, November 30, 2001
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Mimic of Photocycle by a Protein Folding Reaction in Photoactive Yellow Protein*

Byoung-Chul LeeDagger , Paula A. Croonquist, and Wouter D. Hoff§

From the Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, Illinois 60637

The blue light receptor photoactive yellow protein (PYP) displays rhodopsin-like photochemistry based on the trans to cis photoisomerization of its p-coumaric acid chromophore. Here, we report that protein refolding from the acid-denatured state of PYP mimics the last photocycle transition in PYP. This implies a direct link between transient protein unfolding and photosensory signal transduction. We utilize this link to study general issues in protein folding. Chromophore trans to cis photoisomerization in the acid-denatured state strongly decelerates refolding, and converts the pH dependence of the barrier for refolding from linear to nonlinear. We propose transition state movement to explain this phenomenon. The cis chromophore significantly stabilizes the acid-denatured state, but acidification of PYP results in the accumulation of the acid-denatured state containing a trans chromophore. This provides a clear example of kinetic control in a protein unfolding reaction. These results demonstrate the power of PYP as a light-triggered model system to study protein folding.

* This work was supported by American Cancer Society Grant IRG 58-004-41, Cancer Research Foundation Grant 19428, and National Institutes of Health Grant 1 R01 GM63805-01 (all to W. D. H.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Current address: Graduate Group in Biophysics, University of California, San Francisco, CA 94143.

§ To whom correspondence should be addressed. Tel.: 773-834-3098; Fax: 773-702-0439; E-mail: whoff@midway.uchicago.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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