Nuclear magnetic resonance studies of recombinant Escherichia coli glutaredoxin. Sequence-specific assignments and secondary structure determination of the oxidized form.
Escherichia coli glutaredoxin (85 amino acid residues, Mr = 9100), the glutathione-dependent hydrogen donor for ribonucleotide reductase, was purified from an inducible lambda PL, expression system both with a natural isotope content and with uniform 15N labelling. This material was used for obtaining sequence-specific 1H magnetic resonance assignments and the identification of regular secondary structures in the oxidized form of the protein, which contains the redox-active disulfide Cys11-Pro-Tyr-Cys14. Oxidized glutaredoxin contains a four-stranded beta-sheet, with the peripheral strand 32-37 arranged parallel to the strand 2-7, which further combines with the two additional strands 61-64 and 67-69 in an antiparallel fashion. The protein further contains three helices extending approximately from residues 13-28, 45-54 and 72-84.
Sodano
P
P
Institut für Molekularbiologie und Biophysik, Eidgenössiche Technische Hochschule-Hönggerberg, Zürich, Switzerland.
Chary
K V
KV
Björnberg
O
O
Holmgren
A
A
Kren
B
B
Fuchs
J A
JA
Wüthrich
K
K
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D054477:Glutaredoxins; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010088:Oxidoreductases; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry* / Q000235:genetics
D001426:Bacterial Proteins; D054477:Glutaredoxins; D011506:Proteins; D010088:Oxidoreductases