Structural and functional characterization of the mutant Escherichia coli glutaredoxin (C14----S) and its mixed disulfide with glutathione.
Identifieur interne : 001280 ( Main/Exploration ); précédent : 001279; suivant : 001281Structural and functional characterization of the mutant Escherichia coli glutaredoxin (C14----S) and its mixed disulfide with glutathione.
Auteurs : J H Bushweller [Suède] ; F. Aslund ; K. Wüthrich ; A. HolmgrenSource :
- Biochemistry [ 0006-2960 ] ; 1992.
Descripteurs français
- KwdFr :
- Conformation des protéines (MeSH), Disulfures (composition chimique), Disulfures (métabolisme), Données de séquences moléculaires (MeSH), Escherichia coli (génétique), Escherichia coli (métabolisme), Focalisation isoélectrique (MeSH), Glutarédoxines (MeSH), Glutathion (métabolisme), Mutagenèse dirigée (MeSH), Oligodésoxyribonucléotides (MeSH), Oxidoreductases (MeSH), Protéines (composition chimique), Protéines (génétique), Protéines (métabolisme), Protéines bactériennes (composition chimique), Protéines bactériennes (génétique), Protéines bactériennes (métabolisme), Protéines recombinantes (composition chimique), Protéines recombinantes (métabolisme), Spectroscopie par résonance magnétique (MeSH), Séquence d'acides aminés (MeSH), Séquence nucléotidique (MeSH).
- MESH :
- composition chimique : Disulfures, Protéines, Protéines bactériennes, Protéines recombinantes.
- génétique : Escherichia coli, Protéines, Protéines bactériennes.
- métabolisme : Disulfures, Escherichia coli, Glutathion, Protéines, Protéines bactériennes, Protéines recombinantes.
- Conformation des protéines, Données de séquences moléculaires, Focalisation isoélectrique, Glutarédoxines, Mutagenèse dirigée, Oligodésoxyribonucléotides, Oxidoreductases, Spectroscopie par résonance magnétique, Séquence d'acides aminés, Séquence nucléotidique.
English descriptors
- KwdEn :
- Amino Acid Sequence (MeSH), Bacterial Proteins (chemistry), Bacterial Proteins (genetics), Bacterial Proteins (metabolism), Base Sequence (MeSH), Disulfides (chemistry), Disulfides (metabolism), Escherichia coli (genetics), Escherichia coli (metabolism), Glutaredoxins (MeSH), Glutathione (metabolism), Isoelectric Focusing (MeSH), Magnetic Resonance Spectroscopy (MeSH), Molecular Sequence Data (MeSH), Mutagenesis, Site-Directed (MeSH), Oligodeoxyribonucleotides (MeSH), Oxidoreductases (MeSH), Protein Conformation (MeSH), Proteins (chemistry), Proteins (genetics), Proteins (metabolism), Recombinant Proteins (chemistry), Recombinant Proteins (metabolism).
- MESH :
- chemical , chemistry : Bacterial Proteins, Disulfides, Proteins, Recombinant Proteins.
- chemical , genetics : Bacterial Proteins, Proteins.
- chemical , metabolism : Bacterial Proteins, Disulfides, Glutathione, Proteins, Recombinant Proteins.
- genetics : Escherichia coli.
- metabolism : Escherichia coli.
- Amino Acid Sequence, Base Sequence, Glutaredoxins, Isoelectric Focusing, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Mutagenesis, Site-Directed, Oligodeoxyribonucleotides, Oxidoreductases, Protein Conformation.
Abstract
Glutaredoxin is essential for the glutathione (GSH)-dependent synthesis of deoxyribonucleotides by ribonucleotide reductase, and in addition, it displays a general GSH disulfide oxidoreductase activity. In Escherichia coli glutaredoxin, the active site contains a redox-active disulfide/dithiol of the sequence Cys11-Pro12-Tyr13-Cys14. In this paper, we have prepared and characterized the Cys14----Ser mutant of E. coli glutaredoxin and its mixed disulfide with glutathione. The Cys14----Ser mutant of glutaredoxin is shown to retain 38% of the GSH disulfide oxidoreductase activity of the wild-type protein with hydroxyethyl disulfide as substrate but to be completely inactive with ribonucleotide reductase, demonstrating that dithiol glutaredoxin is the hydrogen donor for ribonucleotide reductase. The covalent structure of the mixed disulfide of glutaredoxin(C14S) with GSH prepared with 15N-labeling of the protein was confirmed with nuclear magnetic resonance (NMR) spectroscopy, establishing a basis for NMR structural studies of the glutathione binding site on glutaredoxin.
DOI: 10.1021/bi00153a023
PubMed: 1390715
Affiliations:
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In Escherichia coli glutaredoxin, the active site contains a redox-active disulfide/dithiol of the sequence Cys11-Pro12-Tyr13-Cys14. In this paper, we have prepared and characterized the Cys14----Ser mutant of E. coli glutaredoxin and its mixed disulfide with glutathione. The Cys14----Ser mutant of glutaredoxin is shown to retain 38% of the GSH disulfide oxidoreductase activity of the wild-type protein with hydroxyethyl disulfide as substrate but to be completely inactive with ribonucleotide reductase, demonstrating that dithiol glutaredoxin is the hydrogen donor for ribonucleotide reductase. The covalent structure of the mixed disulfide of glutaredoxin(C14S) with GSH prepared with 15N-labeling of the protein was confirmed with nuclear magnetic resonance (NMR) spectroscopy, establishing a basis for NMR structural studies of the glutathione binding site on glutaredoxin.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">1390715</PMID><DateCompleted><Year>1992</Year><Month>11</Month><Day>13</Day></DateCompleted><DateRevised><Year>2019</Year><Month>06</Month><Day>13</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0006-2960</ISSN><JournalIssue CitedMedium="Print"><Volume>31</Volume><Issue>38</Issue><PubDate><Year>1992</Year><Month>Sep</Month><Day>29</Day></PubDate></JournalIssue><Title>Biochemistry</Title><ISOAbbreviation>Biochemistry</ISOAbbreviation></Journal><ArticleTitle>Structural and functional characterization of the mutant Escherichia coli glutaredoxin (C14----S) and its mixed disulfide with glutathione.</ArticleTitle><Pagination><MedlinePgn>9288-93</MedlinePgn></Pagination><Abstract><AbstractText>Glutaredoxin is essential for the glutathione (GSH)-dependent synthesis of deoxyribonucleotides by ribonucleotide reductase, and in addition, it displays a general GSH disulfide oxidoreductase activity. In Escherichia coli glutaredoxin, the active site contains a redox-active disulfide/dithiol of the sequence Cys11-Pro12-Tyr13-Cys14. In this paper, we have prepared and characterized the Cys14----Ser mutant of E. coli glutaredoxin and its mixed disulfide with glutathione. The Cys14----Ser mutant of glutaredoxin is shown to retain 38% of the GSH disulfide oxidoreductase activity of the wild-type protein with hydroxyethyl disulfide as substrate but to be completely inactive with ribonucleotide reductase, demonstrating that dithiol glutaredoxin is the hydrogen donor for ribonucleotide reductase. The covalent structure of the mixed disulfide of glutaredoxin(C14S) with GSH prepared with 15N-labeling of the protein was confirmed with nuclear magnetic resonance (NMR) spectroscopy, establishing a basis for NMR structural studies of the glutathione binding site on glutaredoxin.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Bushweller</LastName><ForeName>J H</ForeName><Initials>JH</Initials><AffiliationInfo><Affiliation>Department of Biochemistry, Medical Nobel Institute, Karolinska Institute, Stockholm, Sweden.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Aslund</LastName><ForeName>F</ForeName><Initials>F</Initials></Author><Author ValidYN="Y"><LastName>Wüthrich</LastName><ForeName>K</ForeName><Initials>K</Initials></Author><Author ValidYN="Y"><LastName>Holmgren</LastName><ForeName>A</ForeName><Initials>A</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal 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uniqKey="Aslund F" first="F" last="Aslund">F. Aslund</name><name sortKey="Holmgren, A" sort="Holmgren, A" uniqKey="Holmgren A" first="A" last="Holmgren">A. Holmgren</name><name sortKey="Wuthrich, K" sort="Wuthrich, K" uniqKey="Wuthrich K" first="K" last="Wüthrich">K. Wüthrich</name></noCountry><country name="Suède"><region name="Svealand"><name sortKey="Bushweller, J H" sort="Bushweller, J H" uniqKey="Bushweller J" first="J H" last="Bushweller">J H Bushweller</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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