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A designed protein interface that blocks fibril formation

Shukla, Ushma J. and Marino, Heather and Huang, Po-Ssu and Mayo, Stephen L. and Love, John J. (2004) A designed protein interface that blocks fibril formation. Journal of the American Chemical Society, 126 (43). pp. 13914-13915. ISSN 0002-7863. http://resolver.caltech.edu/CaltechAUTHORS:20110913-173424834

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Abstract

Protein fibril formation is implicated in many diseases, and therefore much effort has been focused toward the development of inhibitors of this process. In a previous project, a monomeric protein was computationally engineered to bind itself and form a heterodimer complex following interfacial redesign. One of the protein monomers, termed monomer-B, was unintentionally destabilized and shown to form macroscopic fibrils. Interestingly, in the presence of the designed binding partner, fibril formation was blocked. Here we describe the complete characterization of the amyloid properties of monomer-B and the inhibition of fiber formation by the designed binding partner, monomer-A. Both proteins are mutants of the betal domain of streptococcal protein-G. The free monomer-B protein forms amyloid-type fibrils, as determined by transmission electron microscopy and the change in fluorescence of Thioflavin T, an amyloid-specific dye. Fibril formation kinetics are influenced by pH, protein concentration, and seeding with preformed fibrils. Under all conditions tested, monomer-A was able to inhibit the formation of monomer-B fibrils. This inhibition is specific to the engineered interaction, as incubation of monomer-B with wild-type protein-G (a structural homologue) did not result in inhibition under the same conditions. Thus, this de novo-designed heterodimeric complex is an excellent model system for the study of protein-based fibril formation and inhibition. This system provides additional insight into the development of pharmaceuticals for amyloid disorders, as well as the potential use of amyloid fibrils for self-assembling nanostructures.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/ja0456858 DOIUNSPECIFIED
http://pubs.acs.org/doi/abs/10.1021/ja0456858PublisherUNSPECIFIED
Additional Information:© 2004 American Chemical Society. Received July 19, 2004. Acknowledgment is made for support of this research to the donors of the American Chemical Society Petroleum Research Fund, the Blasker-Rose-Miah fund of the San Diego Foundation, and the California Metabolic Research Foundation.
Funders:
Funding AgencyGrant Number
American Chemical Society Petroleum Research FundUNSPECIFIED
Blasker-Rose-Miah fund of the San Diego FoundationUNSPECIFIED
California Metabolic Research FoundationUNSPECIFIED
Subject Keywords:Amyloid; Bacterial Proteins; Mutagenesis: Site-Directed; Protein Structure: Tertiary; Protein Engineering; Fluorescence; Thiazoles
Record Number:CaltechAUTHORS:20110913-173424834
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20110913-173424834
Official Citation:A Designed Protein Interface That Blocks Fibril Formation Ushma J. Shukla,, Heather Marino,, Po-Ssu Huang,, Stephen L. Mayo, and, John J. Love, Journal of the American Chemical Society 2004 126 (43), 13914-13915
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:25329
Collection:CaltechAUTHORS
Deposited By: Marie Ary
Deposited On:23 Sep 2011 21:20
Last Modified:26 Dec 2012 13:41

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