Mouse glutaredoxin - cDNA cloning, high level expression in E. coli and its possible implication in redox regulation of the DNA binding activity in transcription factor PEBP2.
We have isolated a cDNA encoding glutaredoxin (GRX) from a mouse splenic cDNA library. This cDNA encoded a protein of 107 amino acids with a calculated molecular weight of 11.9 kDa. The deduced amino acid sequence of glutaredoxin in mouse was highly homologous with that in other mammals (81-89%), containing a putative active sequence of -Cys-Pro-Try-Cys-. Recombinant mouse glutaredoxin expressed in E. coli showed glutathione-disulfide oxidoreductase activity with beta-hydroxyethyl disulfide as its substrate, whereas mutant glutaredoxin (Cys 22, Cys 25 to Ser) showed no activity. In electrophoretic mobility shift assay, we proved that wild type GRX, not mutant one, recovered the DNA-binding activity of a transcription factor, PEBP2, oxidized by diamide. This showed that GRX may be involved in the redox regulation of the DNA-binding activity of PEBP2 as is the case with thioredoxin.
Nakamura
T
T
Department of Biological Responses, Institute for Virus Research, Kyoto University, Japan.
Ohno
T
T
Hirota
K
K
Nishiyama
A
A
Nakamura
H
H
Wada
H
H
Yodoi
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli; D054477:Glutaredoxins; D051379:Mice; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010088:Oxidoreductases; D011506:Proteins / Q000235:genetics* / Q000378:metabolism*; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D054477:Glutaredoxins; D011506:Proteins; D050656:Transcription Factor AP-2; D014157:Transcription Factors; D004247:DNA; D010088:Oxidoreductases