| 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 |
9.412
|
| Times Cited |
332
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| 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 |
