Tandem use of selenocysteine

Adaptation of a selenoprotein glutaredoxin for reduction of selenoprotein methionine sulfoxide reductase

Moon Jung Kim, Byung Cheon Lee, Jaeho Jeong, Kong Joo Lee, Kwang Yeon Hwang, Vadim N. Gladyshev, Hwa Young Kim

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Several engineered selenocysteine (Sec)-containing glutaredoxins (Grxs) and their enzymatic properties have been reported, but natural selenoprotein Grxs have not been previously characterized. We expressed a bacterial selenoprotein Grx from Clostridium sp. (also known as Alkaliphilus oremlandii) OhILAs in Escherichia coli and characterized this selenoenzyme and its natural Cys homologues in Clostridium and E. coli. The selenoprotein Grx had a 200-fold higher activity than its Sec-to-Cys mutant form, suggesting that Sec is essential for catalysis by this thiol-disulfide oxidoreductase. Kinetic analysis also showed that the selenoprotein Grx had a 10-fold lower Km than Cys homologues. Interestingly, this selenoenzyme efficiently reduced a Clostridium selenoprotein methionine sulfoxide reductase A (MsrA), suggesting that it is the natural reductant for the protein that is not reducible by thioredoxin, a common reductant for Cys-containing MsrAs. We also found that the selenoprotein Grx could not efficiently reduce a Cys version of Clostridium MsrA, whereas natural Clostridium and E. coli Cys-containing Grxs, which efficiently reduce Cys-containing MsrAs, poorly acted on the selenoprotein MsrA. This specificity for MsrA reduction could explain why Sec is utilized in Clostridium Grx and more generally provides a novel example of the use of Sec in biological systems.

Original languageEnglish
Pages (from-to)1194-1203
Number of pages10
JournalMolecular Microbiology
Volume79
Issue number5
DOIs
Publication statusPublished - 2011 Mar 1

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Glutaredoxins
Selenoproteins
Selenocysteine
Clostridium
Methionine Sulfoxide Reductases
Reducing Agents
Escherichia coli
Protein Disulfide Reductase (Glutathione)
Thioredoxins
methionine sulfoxide reductase
Catalysis

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Tandem use of selenocysteine : Adaptation of a selenoprotein glutaredoxin for reduction of selenoprotein methionine sulfoxide reductase. / Kim, Moon Jung; Lee, Byung Cheon; Jeong, Jaeho; Lee, Kong Joo; Hwang, Kwang Yeon; Gladyshev, Vadim N.; Kim, Hwa Young.

In: Molecular Microbiology, Vol. 79, No. 5, 01.03.2011, p. 1194-1203.

Research output: Contribution to journalArticle

Kim, Moon Jung ; Lee, Byung Cheon ; Jeong, Jaeho ; Lee, Kong Joo ; Hwang, Kwang Yeon ; Gladyshev, Vadim N. ; Kim, Hwa Young. / Tandem use of selenocysteine : Adaptation of a selenoprotein glutaredoxin for reduction of selenoprotein methionine sulfoxide reductase. In: Molecular Microbiology. 2011 ; Vol. 79, No. 5. pp. 1194-1203.
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