A hyperthermostable novel protein-disulfide oxidoreductase is reduced by thioredoxin reductase from hyperthermophilic archaeon Pyrococcus horikoshii.
A redox protein gene (PH0178) with high sequence homology to a glutaredoxin from Pyrococcus furiosus and a thioredoxin reductase homologue gene (PH1426) were found in the genome sequence of Pyrococcus horikoshii. These two genes were cloned and the corresponding expressed proteins were characterized. The redox protein from PH0178 had strong thioredoxin-like activity, but no glutaredoxin activity. The protein from PH1426 had some reductase activity against thioredoxin from Escherichia coli as well as the redox protein (PH0178). The protein from PH1426 was a typical, homodimeric flavoprotein. These results indicate that the redox protein (PH0178) is not a glutaredoxin but, rather, a new protein-disulfide oxidoreductase that is involved in a thioredoxin-like system with thioredoxin reductase (PH1426) in P. horikoshii. The redox protein and thioredoxin reductase retained their full activities for over 1h at 100 degrees C. The redox potential of the redox protein was similar to that of thioredoxin from E. coli and lower than that of glutathione. Site-directed mutagenesis studies revealed that the active site of the redox protein corresponds to a CPYC sequence, located in the middle of the sequence.
Kashima
Yasuhiro
Y
The special division for Human Life Technology, National Institute of Advanced Industrial Science and Technology (AIST Kansai), 1-8-31 Midorigaoka, 563-8577, Ikeda, Osaka, Japan.
Ishikawa
Kazuhiko
K
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004355:Drug Stability; D004789:Enzyme Activation; D004795:Enzyme Stability; D005786:Gene Expression Regulation; D054477:Glutaredoxins; D016297:Mutagenesis, Site-Directed; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry / Q000378:metabolism; D011490:Protein Disulfide Reductase (Glutathione) / Q000737:chemistry* / Q000145:classification / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry* / Q000145:classification / Q000378:metabolism*; D020027:Pyrococcus furiosus / Q000737:chemistry / Q000201:enzymology; D043902:Pyrococcus horikoshii / Q000737:chemistry / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017385:Sequence Homology; D013696:Temperature; D013880:Thioredoxin-Disulfide Reductase / Q000737:chemistry* / Q000378:metabolism*; D013879:Thioredoxins
D054477:Glutaredoxins; D011506:Proteins; D011994:Recombinant Proteins; D013879:Thioredoxins; D010088:Oxidoreductases; D013880:Thioredoxin-Disulfide Reductase; D011490:Protein Disulfide Reductase (Glutathione)