Crystal structure of the Vibrio cholerae colonization factor TcpF and identification of a functional immunogenic site |
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Authors: | Megli Christina J Yuen Alex S W Kolappan Subramaniapillai Richardson Malcolm R Dharmasena Madushini N Krebs Shelly J Taylor Ronald K Craig Lisa |
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Institution: | 1 Department of Microbiology and Immunology, Dartmouth Medical School, Hanover, NH 03755, USA2 Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada V5A 1S6 |
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Abstract: | Vibrio cholerae relies on two main virulence factors—toxin-coregulated pilus (TCP) and cholera toxin—to cause the gastrointestinal disease cholera. TCP is a type IV pilus that mediates bacterial autoagglutination and colonization of the intestine. TCP is encoded by the tcp operon, which also encodes TcpF, a protein of unknown function that is secreted by V. cholerae in a TCP-dependent manner. Although TcpF is not required for TCP biogenesis, a tcpF mutant has a colonization defect in the infant mouse cholera model that is as severe as a pilus mutant. Furthermore, TcpF antisera protect against V. cholerae infection. TcpF has no apparent sequence homology to any known protein. Here, we report the de novo X-ray crystal structure of TcpF and the identification of an epitope that is critical for its function as a colonization factor. A monoclonal antibody recognizing this epitope is protective against V. cholerae challenge and adds to the protection provided by an anti-TcpA antibody. These data suggest that TcpF has a novel function in V. cholerae colonization and define a region crucial for this function. |
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Keywords: | TCP toxin-coregulated pilus mAb monoclonal antibody PBS phosphate-buffered saline NTD N-terminal domain CTD C-terminal domain FnIII fibronectin type III CTLD C-type lectin-like domain EDTA ethylenediaminetetraacetic acid EGTA ethylene glycol bis(β-aminoethyl ether) N N&prime -tetraacetic acid SeMet-TcpF selenomethionine-substituted TcpF ALS Advanced Light Source SSRL Stanford Synchrotron Radiation Laboratory |
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