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Glutaredoxin-3 from Escherichia coli: Amino acid sequence, H-1 and N-15 NMR assignments, and structural analysis
Karolinska Institutet.ORCID iD: 0000-0002-3049-967X
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1996 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 271, no 12, 6736-6745 p.Article in journal (Refereed) PublishedText
Abstract [en]

The primary and secondary structure of glutaredoxin-3 (Grx3), a glutathione-disulfide oxidoreductase from Escherichia coli, has been determined. The amino acid sequence of Grx3 consists of 82 residues and contains a redox-active motif, Cys-Pro-Tyr-Cys, typical of the glutaredoxin family. Sequence comparison reveals a homology (33% identity) to that of glutaredoxin-1 (Grx1) from E. coli as well as to other members of the thioredoxin superfamily. in addition to the active site cysteine residues, Grx3 contains one additional cysteine (Cys(65)) corresponding to one of the two non-active site (or structural) cysteine residues present in mammalian glutaredoxins. The sequence-specific H-1 and N-15 nuclear magnetic resonance assignments of reduced Grx3 have been obtained. From a combined analysis of chemical shifts, (3)J(HN alpha) coupling constants, sequential and medium range NOEs, and amide proton exchange rates, the secondary structure of reduced Grx3 was determined and found to be very similar to that inferred from amino acid sequence comparison to homologous proteins. The consequences of the proposed structural similarity to Grx1 are that Grx3, while possessing a largely intact GSH binding cleft, would have a very different spatial distribution of charged residues, most notably surrounding the active site cysteine residues and occurring in the proposed hydrophobic protein-protein interaction area. These differences may contribute to the observed very low K-cat of Grx3 as a reductant of insulin disulfides or as a hydrogen donor for ribonucleotide reductase. Thus, despite an identical active site disulfide motif and a similar secondary structure and tertiary fold, Grx3 and Grxl display large functional differences in in vitro protein disulfide oxide-reduction reactions.

Place, publisher, year, edition, pages
1996. Vol. 271, no 12, 6736-6745 p.
Keyword [en]
3-dimensional structure, deoxyribonucleotides, glutathione-dependent synthesis, human thioredoxin, mixed disulfide, proteins, proton proton distances, purification, secondary structure determination, spectroscopy
National Category
Structural Biology
Identifiers
URN: urn:nbn:se:sh:diva-28994DOI: 10.1074/jbc.271.12.6736ISI: A1996UB15700029OAI: oai:DiVA.org:sh-28994DiVA: diva2:892178
Note

Times Cited: 28 Article English Cited References Count: 62 Ub157

Available from: 2016-01-08 Created: 2016-01-07 Last updated: 2016-01-11Bibliographically approved

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Berndt, Kurt D
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