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Bio-orthogonal chemistry enables proteomic analysis of Staphylococcus aureus during mammalian infection

Stone, Shannon and Tirrell, David (2016) Bio-orthogonal chemistry enables proteomic analysis of Staphylococcus aureus during mammalian infection. In: 251st American Chemical Society National Meeting & Exposition, March 13-17, 2016, San Diego, CA. https://resolver.caltech.edu/CaltechAUTHORS:20160331-085640195

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Abstract

We report on a new tool to study proteins synthesized by Staphylococcus aureus during infection of a live mouse. Traditional proteomics approaches are often unsuitable to study pathogens in vivo because highly abundant host proteins predominate. To circumvent this issue, we engineered several strains of S. aureus to have the ability to incorporate a noncanonical amino acid with a bio- orthogonal handle. Only these engineered strains of S. aureus can incorporate this amino acid into newly synthesized proteins, thus rendering these proteins chem. distinct from the host proteome, and allowing us to selectively tag and enrich for pathogenic proteins. Before enrichment of proteins tagged in vivo, we found relatively few S. aureus proteins in samples isolated from a skin infection model. After enrichment, we were able to identify 4x more proteins in total, with far greater spectra and sequence coverage per protein. We compared enriched samples from S. aureus labeled in vivo to the proteome synthesized prior to infection (in vitro) . Over 1000 proteins were found in the in vitro lysates, while 484 were identified in the in vivo samples. These results represent the highest no. of S. aureus proteins identified in vivo to date and, in addn. to confirming many previously identified virulence factors, generate new candidates to examine for contributions to virulence.


Item Type:Conference or Workshop Item (Paper)
Related URLs:
URLURL TypeDescription
http://www.acs.org/content/acs/en/meetings/spring-2016.htmlOrganizationConference Website
ORCID:
AuthorORCID
Stone, Shannon0000-0002-6617-3874
Tirrell, David0000-0003-3175-4596
Additional Information:© 2016 American Chemical Society.
Record Number:CaltechAUTHORS:20160331-085640195
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20160331-085640195
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:65787
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:31 Mar 2016 17:49
Last Modified:03 Oct 2019 09:50

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