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Identification of secreted bacterial proteins by noncanonical amino acid tagging

Mahdavi, Alborz and Szychowski, Janek and Ngo, John T. and Sweredoski, Michael J. and Graham, Robert L. J. and Hess, Sonja and Schneewind, Olaf and Mazmanian, Sarkis K. and Tirrell, David A. (2014) Identification of secreted bacterial proteins by noncanonical amino acid tagging. Proceedings of the National Academy of Sciences of the United States of America, 111 (1). pp. 433-438. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:20131224-093455338

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Abstract

Microbial pathogens use complex secretion systems to deliver virulence factors into host cells, where they disrupt host cell function. Understanding these systems is essential to the development of new treatments for infectious disease. A challenge in such studies arises from the abundance of host cell proteins, which interfere with detection of microbial effectors. Here we describe a metabolic labeling strategy that allows selective enrichment of microbial proteins from the host cell cytoplasm. The method enables efficient identification of microbial proteins that have been delivered to the host, identifies distinct secretion profiles for intracellular and extracellular bacteria, and allows for determination of the order of injection of microbial proteins into host cells.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.1301740111DOIArticle
http://www.pnas.org/content/111/1/433PublisherArticle
Additional Information:© 2014 National Academy of Sciences. Edited by Ralph R. Isberg, Howard Hughes Medical Institute, Tufts University School of Medicine, Boston, MA, and approved November 25, 2013 (received for review January 27, 2013). We thank Geoff Smith (Proteome Exploration Laboratory, Beckman Institute, California Institute of Technology) for technical assistance. This work was supported by the National Institutes of Health (Grant R01 GM062523), the Institute for Collaborative Biotechnologies (Grant W911NF-09-0001 from the US Army Research Office), the Burroughs Wellcome Fund in the Pathogenesis of Infectious Disease, and the Gordon and Betty Moore Foundation. A.M. is the recipient of a Natural Sciences and Engineering Reasearch Council of Canada scholarship and a Donna and Benjamin M. Rosen postgraduate scholarship.
Funders:
Funding AgencyGrant Number
NIHR01 GM062523
Institute for Collaborative Biotechnologies (ARO)W911NF-09-0001
Burroughs Wellcome FundUNSPECIFIED
Gordon and Betty Moore FoundationUNSPECIFIED
Natural Sciences and Engineering Reasearch Council of CanadaUNSPECIFIED
Donna and Benjamin M. Rosen postgraduate scholarshipUNSPECIFIED
Subject Keywords:proteomics; click chemistry; BONCAT; Yop
Record Number:CaltechAUTHORS:20131224-093455338
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20131224-093455338
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:43163
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:13 Jan 2014 23:56
Last Modified:14 Nov 2014 19:21

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