期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 106, 期 27, 页码 11067-11072出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0813012106
关键词
beta-sheet nucleator; kinetic assessment of turn mimics; Pin WW domain; protein folding
资金
- Ruth L. Kirschstein National Research Service
- National Institutes of Health (NIH) [GM067082, GM045811, GM057374, GM051105]
- NSF-MCB [0613643]
- Skaggs Institute for Chemical Biology
- Lita Annenberg Hazen Foundation
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [0613643] Funding Source: National Science Foundation
beta-Turns are common conformations that enable proteins to adopt globular structures, and their formation is often rate limiting for folding. beta-Turn mimics, molecules that replace the i + 1 and i + 2 amino acid residues of a beta-turn, are envisioned to act as folding nucleators by preorganizing the pendant polypeptide chains, thereby lowering the activation barrier for beta-sheet formation. However, the crucial kinetic experiments to demonstrate that beta-turn mimics can act as strong nucleators in the context of a cooperatively folding protein have not been reported. We have incorporated 6 beta-turn mimics simulating varied beta-turn types in place of 2 residues in an engineered beta-turn 1 or beta-bulge turn 1 of the Pin 1 WW domain, a three-stranded beta-sheet protein. We present 2 lines of kinetic evidence that the inclusion of beta-turn mimics alters beta-sheet folding rates, enabling us to classify beta-turn mimics into 3 categories: those that are weak nucleators but permit Pin WW folding, native-like nucleators, and strong nucleators. Strong nucleators accelerate folding relative to WW domains incorporating all alpha-amino acid sequences. A solution NMR structure reveals that the native Pin WW beta-sheet structure is retained upon incorporating a strong E-olefin nucleator. These beta-turn mimics can now be used to interrogate protein folding transition state structures and the 2 kinetic analyses presented can be used to assess the nucleation capacity of other beta-turn mimics.
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