Site-directed mutagenesis of glutamic acid 172 to glutamine completely inactivated human O6-alkylguanine-DNA-alkyltransferase.

2.50
Hdl Handle:
http://hdl.handle.net/10541/96106
Title:
Site-directed mutagenesis of glutamic acid 172 to glutamine completely inactivated human O6-alkylguanine-DNA-alkyltransferase.
Authors:
Rafferty, Joseph A; Tumelty, J; Skorvaga, Milan; Elder, Rhoderick H; Margison, Geoffrey P; Douglas, K T
Abstract:
DNA repair by O6-alkylguanine-DNA-alkyltransferase involves the stoichiometric transfer of the O6-alkyl group from the guanine lesion to the active-site cysteine residues of the protein. Site-directed mutagenesis of glutamic acid 172 of human O6-alkylguanine-DNA-alkyltransferase (EC 2.1.1.63) to glutamine totally abolished the alkyltransferase activity of the protein. This suggests that glutamic acid 172 is crucial to the alkyl transfer. It may act as a general acid (as CO2H) or base (as CO2-), or have a role as a component of a salt-link (-CO2-.....+N-), vital for the structural integrity of the active site. This is the first mutational inactivation of a protein in this family of DNA repair molecules by means of a residue change outside the highly conserved pentet (PCHRV) which includes the active-site cysteine.
Affiliation:
CRC Department of Carcinogenesis, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, U.K.
Citation:
Site-directed mutagenesis of glutamic acid 172 to glutamine completely inactivated human O6-alkylguanine-DNA-alkyltransferase. 1994, 199 (1):285-91 Biochem. Biophys. Res. Commun.
Journal:
Biochemical and Biophysical Research Communications
Issue Date:
28-Feb-1994
URI:
http://hdl.handle.net/10541/96106
DOI:
10.1006/bbrc.1994.1226
PubMed ID:
8123025
Type:
Article
Language:
en
ISSN:
0006-291X
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorRafferty, Joseph Aen
dc.contributor.authorTumelty, Jen
dc.contributor.authorSkorvaga, Milanen
dc.contributor.authorElder, Rhoderick Hen
dc.contributor.authorMargison, Geoffrey Pen
dc.contributor.authorDouglas, K Ten
dc.date.accessioned2010-04-09T09:22:24Z-
dc.date.available2010-04-09T09:22:24Z-
dc.date.issued1994-02-28-
dc.identifier.citationSite-directed mutagenesis of glutamic acid 172 to glutamine completely inactivated human O6-alkylguanine-DNA-alkyltransferase. 1994, 199 (1):285-91 Biochem. Biophys. Res. Commun.en
dc.identifier.issn0006-291X-
dc.identifier.pmid8123025-
dc.identifier.doi10.1006/bbrc.1994.1226-
dc.identifier.urihttp://hdl.handle.net/10541/96106-
dc.description.abstractDNA repair by O6-alkylguanine-DNA-alkyltransferase involves the stoichiometric transfer of the O6-alkyl group from the guanine lesion to the active-site cysteine residues of the protein. Site-directed mutagenesis of glutamic acid 172 of human O6-alkylguanine-DNA-alkyltransferase (EC 2.1.1.63) to glutamine totally abolished the alkyltransferase activity of the protein. This suggests that glutamic acid 172 is crucial to the alkyl transfer. It may act as a general acid (as CO2H) or base (as CO2-), or have a role as a component of a salt-link (-CO2-.....+N-), vital for the structural integrity of the active site. This is the first mutational inactivation of a protein in this family of DNA repair molecules by means of a residue change outside the highly conserved pentet (PCHRV) which includes the active-site cysteine.en
dc.language.isoenen
dc.subject.meshAmino Acid Sequence-
dc.subject.meshDNA Repair-
dc.subject.meshGlutamates-
dc.subject.meshGlutamine-
dc.subject.meshHumans-
dc.subject.meshMethyltransferases-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshMutagenesis, Site-Directed-
dc.subject.meshO(6)-Methylguanine-DNA Methyltransferase-
dc.subject.meshStructure-Activity Relationship-
dc.titleSite-directed mutagenesis of glutamic acid 172 to glutamine completely inactivated human O6-alkylguanine-DNA-alkyltransferase.en
dc.typeArticleen
dc.contributor.departmentCRC Department of Carcinogenesis, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, U.K.en
dc.identifier.journalBiochemical and Biophysical Research Communicationsen
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