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dc.contributor.authorElder, Rhoderick H
dc.contributor.authorTumelty, J
dc.contributor.authorDouglas, K T
dc.contributor.authorMargison, Geoffrey P
dc.contributor.authorRafferty, Joseph A
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.
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
html.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.


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