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A de novo designed protein-protein interface

Huang, Po-Ssu and Love, John J and Mayo, Stephen L. (2007) A de novo designed protein-protein interface. Protein Science, 16 (12). pp. 2770-2774. ISSN 0961-8368. http://resolver.caltech.edu/CaltechAUTHORS:20110913-160000725

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Abstract

As an approach to both explore the physical/chemical parameters that drive molecular self-assembly and to generate novel protein oligomers, we have developed a procedure to generate protein dimers from monomeric proteins using computational protein docking and amino acid sequence design. A fast Fourier transform-based docking algorithm was used to generate a model for a dimeric version of the 56-amino-acid β1 domain of streptococcal protein G. Computational amino acid sequence design of 24 residues at the dimer interface resulted in a heterodimer comprised of 12-fold and eightfold variants of the wild-type protein. The designed proteins were expressed, purified, and characterized using analytical ultracentrifugation and heteronuclear NMR techniques. Although the measured dissociation constant was modest (~300 µM), 2D-[^1H,^(15)N]-HSQC NMR spectra of one of the designed proteins in the absence and presence of its binding partner showed clear evidence of specific dimer formation.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1110/ps.073125207DOIUNSPECIFIED
http://onlinelibrary.wiley.com/doi/10.1110/ps.073125207/abstract;jsessionid=00C4D629513CDD6CCD973A9A3CFA4AC7.d01t01PublisherUNSPECIFIED
Contact Email Address:steve@mayo.caltech.edu
Additional Information:© 2007 The Protein Society. Published by Cold Spring Harbor Laboratory Press. Received July 16, 2007; Final Revision September 21, 2007; Accepted September 21, 2007. This work was supported by the Howard Hughes Medical Institute, the Ralph M. Parsons Foundation, DARPA, the Institute for Collaborative Biotechnologies through grant DAAD19-03-D-0004 from the U.S. Army Research Office, and an IBM Shared University Research Grant.
Funders:
Funding AgencyGrant Number
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Ralph M. Parsons FoundationUNSPECIFIED
Defense Advanced Research Projects Agency (DARPA)UNSPECIFIED
Institute for Collaborative BiotechnologiesDAAD19-03-D-0004
IBM Shared University Research GrantUNSPECIFIED
Subject Keywords:de novo protein–protein interface; computational protein design; geometric recognition algorithm; protein G; heterodimer; NMR; docking
Record Number:CaltechAUTHORS:20110913-160000725
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20110913-160000725
Official Citation:Huang, P.-S., Love, J. J. and Mayo, S. L. (2007), A de novo designed protein–protein interface. Protein Science, 16: 2770–2774. doi: 10.1110/ps.073125207
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:25321
Collection:CaltechAUTHORS
Deposited By: Marie Ary
Deposited On:15 Sep 2011 18:09
Last Modified:23 Aug 2016 10:05

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