PMID:15543158
| Citation |
Lee, C, Lee, SM, Mukhopadhyay, P, Kim, SJ, Lee, SC, Ahn, WS, Yu, MH, Storz, G and Ryu, SE (2004) Redox regulation of OxyR requires specific disulfide bond formation involving a rapid kinetic reaction path. Nat. Struct. Mol. Biol. 11:1179-85 |
|---|---|
| Abstract |
The Escherichia coli OxyR transcription factor is activated by cellular hydrogen peroxide through the oxidation of reactive cysteines. Although there is substantial evidence for specific disulfide bond formation in the oxidative activation of OxyR, the presence of the disulfide bond has remained controversial. By mass spectrometry analyses and in vivo labeling assays we found that oxidation of OxyR in the formation of a specific disulfide bond between Cys199 and Cys208 in the wild-type protein. In addition, using time-resolved kinetic analyses, we determined that OxyR activation occurs at a rate of 9.7 s(-1). The disulfide bond-mediated conformation switch results in a metastable form that is locally strained by approximately 3 kcal mol(-1). On the basis of these observations we conclude that OxyR activation requires specific disulfide bond formation and that the rapid kinetic reaction path and conformation strain, respectively, drive the oxidation and reduction of OxyR. |
| Links |
PubMed Online version:10.1038/nsmb856 |
| Keywords |
DNA-Binding Proteins/chemistry; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism; Disulfides/chemistry; Disulfides/metabolism; Escherichia coli/chemistry; Escherichia coli/drug effects; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli Proteins; Kinetics; Models, Molecular; Oxidants/pharmacology; Oxidation-Reduction/drug effects; Protein Denaturation/drug effects; Protein Folding; Protein Structure, Tertiary; Repressor Proteins/chemistry; Repressor Proteins/genetics; Repressor Proteins/metabolism; Spectrometry, Fluorescence; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Thermodynamics; Transcription Factors/chemistry; Transcription Factors/genetics; Transcription Factors/metabolism; Tryptophan/metabolism; Urea/pharmacology |
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