Site directed mutagenesis of two cysteine residues in the E. coli ogt O6-alkylguanine DNA alkyltransferase protein.

2.50
Hdl Handle:
http://hdl.handle.net/10541/108917
Title:
Site directed mutagenesis of two cysteine residues in the E. coli ogt O6-alkylguanine DNA alkyltransferase protein.
Authors:
Harris, L C; Potter, P M; Margison, Geoffrey P
Abstract:
The E. coli ogt O6-alkylguanine-DNA alkyltransferase has two cysteine residues positioned identically with respect to cysteines in the E. coli ada O6-alkylguanine-DNA alkyltransferase. In order to assess their function, these residues were each substituted by a glycine to generate altered forms of the ogt protein. Mutagenesis of cysteine-139, located within a 'PCHRV' region of homology, eliminated functional activity confirming that this residue is the methyl-accepting cysteine in the active site of the protein. Substitution of cysteine 102 within the sequence 'LRTIPCG' had little effect on the ogt protein activity demonstrating that this cysteine is not directly involved with the transfer of O6-methylguanine adducts.
Affiliation:
CRC Department of Chemical Carcinogenesis, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Manchester, UK.
Citation:
Site directed mutagenesis of two cysteine residues in the E. coli ogt O6-alkylguanine DNA alkyltransferase protein. 1992, 187 (1):425-31 Biochem. Biophys. Res. Commun.
Journal:
Biochemical and Biophysical Research Communications
Issue Date:
31-Aug-1992
URI:
http://hdl.handle.net/10541/108917
DOI:
10.1016/S0006-291X(05)81510-4
PubMed ID:
1520330
Type:
Article
Language:
en
ISSN:
0006-291X
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorHarris, L Cen
dc.contributor.authorPotter, P Men
dc.contributor.authorMargison, Geoffrey Pen
dc.date.accessioned2010-08-03T10:48:40Z-
dc.date.available2010-08-03T10:48:40Z-
dc.date.issued1992-08-31-
dc.identifier.citationSite directed mutagenesis of two cysteine residues in the E. coli ogt O6-alkylguanine DNA alkyltransferase protein. 1992, 187 (1):425-31 Biochem. Biophys. Res. Commun.en
dc.identifier.issn0006-291X-
dc.identifier.pmid1520330-
dc.identifier.doi10.1016/S0006-291X(05)81510-4-
dc.identifier.urihttp://hdl.handle.net/10541/108917-
dc.description.abstractThe E. coli ogt O6-alkylguanine-DNA alkyltransferase has two cysteine residues positioned identically with respect to cysteines in the E. coli ada O6-alkylguanine-DNA alkyltransferase. In order to assess their function, these residues were each substituted by a glycine to generate altered forms of the ogt protein. Mutagenesis of cysteine-139, located within a 'PCHRV' region of homology, eliminated functional activity confirming that this residue is the methyl-accepting cysteine in the active site of the protein. Substitution of cysteine 102 within the sequence 'LRTIPCG' had little effect on the ogt protein activity demonstrating that this cysteine is not directly involved with the transfer of O6-methylguanine adducts.en
dc.language.isoenen
dc.subject.meshAmino Acid Sequence-
dc.subject.meshBase Sequence-
dc.subject.meshBinding Sites-
dc.subject.meshBlotting, Western-
dc.subject.meshCysteine-
dc.subject.meshDNA, Recombinant-
dc.subject.meshEscherichia coli-
dc.subject.meshKinetics-
dc.subject.meshMethyltransferases-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshMutagenesis, Site-Directed-
dc.subject.meshO(6)-Methylguanine-DNA Methyltransferase-
dc.subject.meshRecombinant Fusion Proteins-
dc.subject.meshStructure-Activity Relationship-
dc.titleSite directed mutagenesis of two cysteine residues in the E. coli ogt O6-alkylguanine DNA alkyltransferase protein.en
dc.typeArticleen
dc.contributor.departmentCRC Department of Chemical Carcinogenesis, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Manchester, UK.en
dc.identifier.journalBiochemical and Biophysical Research Communicationsen
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