C-terminally truncated human O6-alkylguanine-DNA alkyltransferase retains activity.

2.50
Hdl Handle:
http://hdl.handle.net/10541/108927
Title:
C-terminally truncated human O6-alkylguanine-DNA alkyltransferase retains activity.
Authors:
Elder, Rhoderick H; Tumelty, J; Douglas, K T; Margison, Geoffrey P; Rafferty, Joseph A
Abstract:
A cDNA encoding the human O6-alkylguanine-DNA alkyltransferase (ATase; EC 2.1.1.63; methylated-DNA: protein-cysteine methyltransferase) has been manipulated to generate a C-terminally deleted protein which retains full methyl-transfer activity. The elimination of 22 amino-acid residues from the C-terminus was achieved by endonuclease-SacI digestion of the 623 bp cDNA coding sequence and ligation of a SacI/HindIII linker containing an in-frame stop codon. The truncated protein was characterized by its reduced molecular mass in immunoblots probed with an antiserum against the full-length protein and by fluorography after incubation with [3H]methylated calf thymus DNA. The rate of methyl transfer was virtually identical for the full-length and truncated ATases. The construction of such a truncated, yet still functional, ATase, with a molecular mass of 19.7 kDa should facilitate a detailed n.m.r. structural study and help to determine the functional significance of the C-terminal domain of mammalian ATases.
Affiliation:
CRC Department of Carcinogenesis, Paterson Institute for Cancer Research, Christie Hospital (NHS) Trust, Manchester, U.K.
Citation:
C-terminally truncated human O6-alkylguanine-DNA alkyltransferase retains activity. 1992, 285 ( Pt 3):707-9 Biochem. J.
Journal:
Biochemical Journal
Issue Date:
1-Aug-1992
URI:
http://hdl.handle.net/10541/108927
PubMed ID:
1497608
Type:
Article
Language:
en
ISSN:
0264-6021
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorElder, Rhoderick Hen
dc.contributor.authorTumelty, Jen
dc.contributor.authorDouglas, K Ten
dc.contributor.authorMargison, Geoffrey Pen
dc.contributor.authorRafferty, Joseph Aen
dc.date.accessioned2010-08-03T11:39:12Z-
dc.date.available2010-08-03T11:39:12Z-
dc.date.issued1992-08-01-
dc.identifier.citationC-terminally truncated human O6-alkylguanine-DNA alkyltransferase retains activity. 1992, 285 ( Pt 3):707-9 Biochem. J.en
dc.identifier.issn0264-6021-
dc.identifier.pmid1497608-
dc.identifier.urihttp://hdl.handle.net/10541/108927-
dc.description.abstractA cDNA encoding the human O6-alkylguanine-DNA alkyltransferase (ATase; EC 2.1.1.63; methylated-DNA: protein-cysteine methyltransferase) has been manipulated to generate a C-terminally deleted protein which retains full methyl-transfer activity. The elimination of 22 amino-acid residues from the C-terminus was achieved by endonuclease-SacI digestion of the 623 bp cDNA coding sequence and ligation of a SacI/HindIII linker containing an in-frame stop codon. The truncated protein was characterized by its reduced molecular mass in immunoblots probed with an antiserum against the full-length protein and by fluorography after incubation with [3H]methylated calf thymus DNA. The rate of methyl transfer was virtually identical for the full-length and truncated ATases. The construction of such a truncated, yet still functional, ATase, with a molecular mass of 19.7 kDa should facilitate a detailed n.m.r. structural study and help to determine the functional significance of the C-terminal domain of mammalian ATases.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshCattle-
dc.subject.meshCloning, Molecular-
dc.subject.meshDNA-
dc.subject.meshDeoxyribonucleases, Type II Site-Specific-
dc.subject.meshEscherichia coli-
dc.subject.meshHumans-
dc.subject.meshImmunoblotting-
dc.subject.meshKinetics-
dc.subject.meshMethylation-
dc.subject.meshMethylnitrosourea-
dc.subject.meshMethyltransferases-
dc.subject.meshMolecular Weight-
dc.subject.meshO(6)-Methylguanine-DNA Methyltransferase-
dc.subject.meshPeptide Fragments-
dc.subject.meshStructure-Activity Relationship-
dc.subject.meshTransformation, Bacterial-
dc.titleC-terminally truncated human O6-alkylguanine-DNA alkyltransferase retains activity.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 Journalen
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