van Aalten, D. M., Haker, A., Hendriks, J., Hellingwerf, K. J., Joshua-Tor, L., Crielaard, W. (February 2002) Engineering photocycle dynamics. Crystal structures and kinetics of three photoactive yellow protein hinge-bending mutants. J Biol Chem, 277 (8). pp. 6463-8. ISSN 0021-9258 (Print) (Public Dataset)
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Abstract
Crystallographic and spectroscopic analyses of three hinge-bending mutants of the photoactive yellow protein are described. Previous studies have identified Gly(47) and Gly(51) as possible hinge points in the structure of the protein, allowing backbone segments around the chromophore to undergo large concerted motions. We have designed, crystallized, and solved the structures of three mutants: G47S, G51S, and G47S/G51S. The protein dynamics of these mutants are significantly affected. Transitions in the photocycle, measured with laser induced transient absorption spectroscopy, show rates up to 6-fold different from the wild type protein and show an additive effect in the double mutant. Compared with the native structure, no significant conformational differences were observed in the structures of the mutant proteins. We conclude that the structural and dynamic integrity of the region around these mutations is of crucial importance to the photocycle and suggest that the hinge-bending properties of Gly(51) may also play a role in PAS domain proteins where it is one of the few conserved residues.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Amino Acid Sequence Amino Acid Substitution Bacterial Proteins chemistry genetics metabolism Crystallography, X-Ray Escherichia coli Gammaproteobacteria metabolism radiation effects Kinetics Light Models Molecular Mutagenesis Site-Directed Photoreceptors Microbial Protein Conformation |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein structure rendering Investigative techniques and equipment > x ray crystallography |
| CSHL Authors: | |
| Communities: | CSHL labs > Joshua-Tor lab |
| Depositing User: | CSHL Librarian |
| Date: | 22 February 2002 |
| Date Deposited: | 21 Mar 2012 19:51 |
| Last Modified: | 05 Sep 2017 19:12 |
| Related URLs: | |
| Dataset ID: | |
| URI: | https://repository.cshl.edu/id/eprint/25489 |
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