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Generation and analysis of proline mutants in protein G

Choi, Eun Jung and Mayo, Stephen L (2006) Generation and analysis of proline mutants in protein G. Protein Engineering, Design and Selection, 19 (6). pp. 285-289. ISSN 1741-0126. doi:10.1093/protein/gzl007.

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The pyrrolidine ring of the amino acid proline reduces the conformational freedom of the protein backbone in its unfolded form and thus enhances protein stability. The strategy of inserting proline into regions of the protein where it does not perturb the structure has been utilized to stabilize many different proteins including enzymes. However, most of these efforts have been based on trial and error, rather than rational design. Here, we try to understand proline's effect on protein stability by introducing proline mutations into various regions of the B1 domain of Streptococcal protein G. We also applied the Optimization of Rotamers By Iterative Techniques computational protein design program, using two different solvation models, to determine the extent to which it could predict the stabilizing and destabilizing effects of prolines. Use of a surface area dependent solvation model resulted in a modest correlation between the experimental free energy of folding and computed energies; on the other hand, use of a Gaussian solvent exclusion model led to significant positive correlation. Including a backbone conformational entropy term to the computational energies increases the statistical significance of the correlation between the experimental stabilities and both solvation models.

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Mayo, Stephen L0000-0002-9785-5018
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Additional Information:© The Author 2006. Published by Oxford University Press. Received February 16, 2006; accepted February 17, 2006. The authors thank I. Caglar Tanrikulu for the helpful discussions and Marie Ary for editing the manuscript. This work was supported by the Howard Hughes Medical Institute, the Ralph M. Parsons Foundation and an IBM Shared University Research Grant.
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Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Ralph M. Parsons FoundationUNSPECIFIED
Subject Keywords:proline, protein, design, protein G, protein stability
Issue or Number:6
Record Number:CaltechAUTHORS:20110913-165232835
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:25328
Deposited By: Marie Ary
Deposited On:15 Sep 2011 17:24
Last Modified:09 Nov 2021 16:32

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