Direct detection of bacterial protein secretion using whole colony proteomics.

Molecular & Cellular Proteomics : MCP
Matthew M ChampionPatricia A DiGiuseppe Champion

Abstract

Bacteria use a variety of secretion systems to transport proteins beyond their cell membrane to interact with their environment. For bacterial pathogens, these systems are key virulence determinants that transport bacterial proteins into host cells. Genetic screens to identify bacterial genes required for export have relied on enzymatic or fluorescent reporters fused to known substrates to monitor secretion. However, they cannot be used in analysis of all secretion systems, limiting the implementation across bacteria. Here, we introduce the first application of a modified form of whole colony MALDI-TOF MS to directly detect protein secretion from intact bacterial colonies. We show that this method is able to specifically monitor the ESX-1 system protein secretion system, a major virulence determinant in both mycobacterial and Gram-positive pathogens that is refractory to reporter analysis. We validate the use of this technology as a high throughput screening tool by identifying an ESAT-6 system 1-deficient mutant from a Mycobacterium marinum transposon insertion library. Furthermore, we also demonstrate detection of secreted proteins of the prevalent type III secretion system from the Gram-negative pathogen, Pseudomonas aerugin...Continue Reading

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Citations

Mar 27, 2013·Applied Microbiology and Biotechnology·Kheng Oon LowRosli Md Illias
May 8, 2014·MBio·M Sloan SiegristEric J Rubin
Oct 15, 2013·Journal of Bacteriology·Patricia A DiGiuseppe Champion
Mar 13, 2014·The American Journal of Tropical Medicine and Hygiene·Nicholas S GeraciMary Ann McDowell
Dec 1, 2012·Expert Review of Proteomics·Jean ArmengaudCatherine Duport
Apr 29, 2015·The Analyst·Sukhendu NandiRaz Jelinek
Aug 12, 2014·Journal of Proteome Research·Matthew M ChampionPatricia A DiGiuseppe Champion
May 8, 2016·Biology of Blood and Marrow Transplantation : Journal of the American Society for Blood and Marrow Transplantation·Maneesh BhargavaChris H Wendt
Aug 3, 2012·Journal of Laboratory Automation·Armand G Ngounou WetieCostel C Darie
Feb 7, 2017·Molecular & Cellular Proteomics : MCP·Todd M Greco, Ileana M Cristea
Nov 1, 2016·Infection and Immunity·Emily A WilliamsPatricia A Champion
May 4, 2017·Journal of Bacteriology·Rachel E Bosserman, Patricia A Champion
Mar 13, 2014·Journal of Bacteriology·George M KennedyPatricia A DiGiuseppe Champion
Aug 20, 2014·Infection and Immunity·Felix Mba MediePatricia A DiGiuseppe Champion
Mar 6, 2019·Journal of Bacteriology·Rachel E BossermanPatricia A Champion
Sep 4, 2020·Microbiology and Molecular Biology Reviews : MMBR·Alexandra E ChirakosPatricia A Champion
May 8, 2014·Molecular Cancer Research : MCR·Kelsey J WeigelZachary T Schafer
Jan 8, 2021·PLoS Pathogens·Kathleen R NicholsonPatricia A Champion
Aug 7, 2018·Journal of Proteome Research·Cristal Reyna ThompsonPatricia A Champion

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Methods Mentioned

BETA
acetylation
Assay

Software Mentioned

Marinolist
LI
Flex Analysis
ProtScore
- Odyssey
ABSciex
Protein Pilot ( ABSciex )
BLAST

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