RRC ID 4929
Author Leichert LI, Gehrke F, Gudiseva HV, Blackwell T, Ilbert M, Walker AK, Strahler JR, Andrews PC, Jakob U.
Title Quantifying changes in the thiol redox proteome upon oxidative stress in vivo.
Journal Proc. Natl. Acad. Sci. U.S.A.
Abstract Antimicrobial levels of reactive oxygen species (ROS) are produced by the mammalian host defense to kill invading bacteria and limit bacterial colonization. One main in vivo target of ROS is the thiol group of proteins. We have developed a quantitative thiol trapping technique termed OxICAT to identify physiologically important target proteins of hydrogen peroxide (H(2)O(2)) and hypochlorite (NaOCl) stress in vivo. OxICAT allows the precise quantification of oxidative thiol modifications in hundreds of different proteins in a single experiment. It also identifies the affected proteins and defines their redox-sensitive cysteine(s). Using this technique, we identified a group of Escherichia coli proteins with significantly (30-90%) oxidatively modified thiol groups, which appear to be specifically sensitive to either H(2)O(2) or NaOCl stress. These results indicate that individual oxidants target distinct proteins in vivo. Conditionally essential E. coli genes encode one-third of redox-sensitive proteins, a finding that might explain the bacteriostatic effect of oxidative stress treatment. We identified a select group of redox-regulated proteins, which protect E. coli against oxidative stress conditions. These experiments illustrate that OxICAT, which can be used in a variety of different cell types and organisms, is a powerful tool to identify, quantify, and monitor oxidative thiol modifications in vivo.
Volume 105(24)
Pages 8197-202
Published 2008-6-17
DOI 10.1073/pnas.0707723105
PII 0707723105
PMID 18287020
PMC PMC2448814
MeSH Animals Escherichia coli / drug effects Escherichia coli / genetics Escherichia coli / metabolism* Escherichia coli Proteins / chemistry Escherichia coli Proteins / genetics Escherichia coli Proteins / metabolism* Hydrogen Peroxide / pharmacology Hypochlorous Acid / pharmacology Mass Spectrometry / methods Oxidation-Reduction Oxidative Stress* / genetics Proteome* Reactive Oxygen Species / metabolism* Sulfhydryl Compounds / analysis Sulfhydryl Compounds / metabolism*
IF 9.58
Times Cited 245
Prokaryotes E. coli ME9062(BW25113) JW2929 JW0213 JW3907 JW2535 JW5300 JW3205 JW1317 JW3585 JW2667 JW2011 JW0598 JW3475 JW3301 JW1104 JW4217 JW2943 JW2491 JW2502 JW0002 JW3928 JW1228