Mechanism of 1-Cys type methionine sulfoxide reductase A regeneration by glutaredoxin

Moon Jung Kim, Jaeho Jeong, Jihye Jeong, Kwang Yeon Hwang, Kong Joo Lee, Hwa Young Kim

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Glutaredoxin (Grx), a major redox regulator, can act as a reductant of methionine sulfoxide reductase A (MsrA). However, the biochemical mechanisms involved in MsrA activity regeneration by Grx remain largely unknown. In this study, we investigated the regeneration mechanism of 1-Cys type Clostridium oremlandii MsrA (cMsrA) lacking a resolving Cys residue in a Grx-dependent assay. Kinetic analysis showed that cMsrA could be reduced by both monothiol and dithiol Grxs as efficiently as by in vitro reductant dithiothreitol. Our data revealed that the catalytic Cys sulfenic acid intermediate is not glutathionylated in the presence of the substrate, and that Grx instead directly formed a complex with cMsrA. Mass spectrometry analysis identified a disulfide bond between the N-terminal catalytic Cys of the active site of Grx and the catalytic Cys of cMsrA. This mixed disulfide bond could be resolved by glutathione. Based on these findings, we propose a model for regeneration of 1-Cys type cMsrA by Grx that involves no glutathionylation on the catalytic Cys of cMsrA. This mechanism contrasts with that of the previously known 1-Cys type MsrB.

Original languageEnglish
Pages (from-to)567-571
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume457
Issue number4
DOIs
Publication statusPublished - 2015 Feb 20

Fingerprint

Methionine Sulfoxide Reductases
Glutaredoxins
Clostridium
Regeneration
Reducing Agents
Disulfides
Dithiothreitol
Oxidation-Reduction
Mass spectrometry
Glutathione
Assays
Mass Spectrometry
Catalytic Domain
Kinetics
Substrates

Keywords

  • 1-Cys
  • Disulfide bond
  • Glutaredoxin
  • Glutathionylation
  • MsrA
  • Regeneration

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Mechanism of 1-Cys type methionine sulfoxide reductase A regeneration by glutaredoxin. / Kim, Moon Jung; Jeong, Jaeho; Jeong, Jihye; Hwang, Kwang Yeon; Lee, Kong Joo; Kim, Hwa Young.

In: Biochemical and Biophysical Research Communications, Vol. 457, No. 4, 20.02.2015, p. 567-571.

Research output: Contribution to journalArticle

Kim, Moon Jung ; Jeong, Jaeho ; Jeong, Jihye ; Hwang, Kwang Yeon ; Lee, Kong Joo ; Kim, Hwa Young. / Mechanism of 1-Cys type methionine sulfoxide reductase A regeneration by glutaredoxin. In: Biochemical and Biophysical Research Communications. 2015 ; Vol. 457, No. 4. pp. 567-571.
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