Linked Life Data

9111025

Source:http://linkedlifedata.com/resource/pubmed/id/9111025

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
17
pubmed:dateCreated
1997-5-21
pubmed:databankReference
pubmed:abstractText
Glutaredoxin 2 (Grx2) from Escherichia coli catalyzes GSH-disulfide oxidoreductions via two redox-active cysteine residues, but in contrast to glutaredoxin 1 (Grx1) and glutaredoxin 3 (Grx3), is not a hydrogen donor for ribonucleotide reductase. To characterize Grx2, a chromosomal fragment containing the E. coli Grx2 gene (grxB) was cloned and sequenced. grxB (645 base pairs) is located between the rimJ and pyrC genes while an open reading frame immediately upstream grxB encodes a novel transmembrane protein of 402 amino acids potentially belonging to class II of substrate export transporters. The deduced amino acid sequence for Grx2 comprises 215 residues with a molecular mass of 24.3 kDa. There is almost no similarity between the amino acid sequence of Grx2 and Grx1 or Grx3 (both 9-kDa proteins) with the exception of the active site which is identical in all three glutaredoxins (C9PYC12 for Grx2). Only limited similarities were noted to glutathione S-transferases (Grx2 amino acids 16-72), and protein disulfide isomerases from different organisms (Grx2 amino acids 70-180). Grx2 was overexpressed and purified to homogeneity and its activity was compared with those of Grx1 and Grx3 using GSH, NADPH, and glutathione reductase in the reduction of 0.7 mM beta-hydroxyethyl disulfide. The three glutaredoxins had similar apparent Km values for GSH (2-3 mM) but Grx2 had the highest apparent kcat (554 s-1). Expression of two truncated forms of Grx2 (1-114 and 1-133) which have predicted secondary structures similar to Grx1 (betaalphabetaalphabetabetaalpha) gave rise to inclusion bodies. The mutant proteins were resolubilized and purified but lacked GSH-disulfide oxidoreductase activity. The latter should therefore require the participation of amino acid residues from the COOH-terminal half of the molecule and is probably not confined to a Grx1-like NH2-terminal subdomain. Grx2 being radically different from the presently known glutaredoxins in terms of molecular weight, amino acid sequence, catalytic activity, and lack of a consensus GSH-binding site is the first member of a novel class of glutaredoxins.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
25
pubmed:volume
272
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
11236-43
pubmed:dateRevised
2011-11-17
pubmed:meshHeading
pubmed-meshheading:9111025-Amino Acid Sequence, pubmed-meshheading:9111025-Bacterial Proteins, pubmed-meshheading:9111025-Base Sequence, pubmed-meshheading:9111025-Cloning, Molecular, pubmed-meshheading:9111025-Escherichia coli, pubmed-meshheading:9111025-Genes, Bacterial, pubmed-meshheading:9111025-Glutaredoxins, pubmed-meshheading:9111025-Glutathione, pubmed-meshheading:9111025-Insulin, pubmed-meshheading:9111025-Membrane Proteins, pubmed-meshheading:9111025-Models, Molecular, pubmed-meshheading:9111025-Molecular Sequence Data, pubmed-meshheading:9111025-Mutation, pubmed-meshheading:9111025-Oxidation-Reduction, pubmed-meshheading:9111025-Oxidoreductases, pubmed-meshheading:9111025-Protein Biosynthesis, pubmed-meshheading:9111025-Protein Structure, Secondary, pubmed-meshheading:9111025-Proteins, pubmed-meshheading:9111025-Recombinant Proteins, pubmed-meshheading:9111025-Restriction Mapping, pubmed-meshheading:9111025-Sequence Analysis, DNA, pubmed-meshheading:9111025-Sequence Deletion, pubmed-meshheading:9111025-Sequence Homology, Amino Acid, pubmed-meshheading:9111025-Species Specificity
pubmed:year
1997
pubmed:articleTitle
Cloning, overexpression, and characterization of glutaredoxin 2, an atypical glutaredoxin from Escherichia coli.
pubmed:affiliation
Medical Nobel Institute for Biochemistry, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm, Sweden.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't