WILKINSON, MC;
POTTER, PM;
CAWKWELL, L;
GEORGIADIS, P;
PATEL, D;
SWANN, PF;
MARGISON, GP;
(1989)
Purification of the Escherichia-coli OGT gene-product to homogeneity and its rate of action on O6-methylguanine, O6-ethylguanine and O4-methylthymine in dodecadeoxyribnucleotides.
Nucleic Acids Research
, 17
(21)
8475 - 8484.
10.1093/nar/17.21.8475.
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Abstract
The E.coli gene ogt encodes the DNA repair protein O6-alkylguanine-DNA-alkyltransferase (O6-AlkG ATase). The protein coding region of the gene was cloned into a multicopy expression vector to obtain high yields of the enzyme (˜ 0.2% of total protein) which was purified to apparent homogeneity by affinity, molecular exclusion and reverse-phase chromatography. Good correlation was found between the determined and predicted amino acid compositions. The ability of the purified protein to act on O6-methylguanine (O6-MeG), O6-ethylguanine (O6-EtG) and (O4-methylthymine (O4-MeT) in self-complementary dodecadeoxyribonucleotides was compared to that of 19 kDa fragment of the related ada-protein. With both proteins the rate order was O6-MeG > O6-EtG > O4-MeT, however, the ogt protein was found to repair O6MeG, O6-EtG and (O4-MeT, 1.1, 173 and 84 times, respectively, faster than the ada protein.
| Type: | Article |
|---|---|
| Title: | Purification of the Escherichia-coli OGT gene-product to homogeneity and its rate of action on O6-methylguanine, O6-ethylguanine and O4-methylthymine in dodecadeoxyribnucleotides |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1093/nar/17.21.8475 |
| Publisher version: | http://dx.doi.org/10.1093/nar/17.21.8475 |
| Language: | English |
| Additional information: | This work is published under the Open Access terms of the Nucleic Acids Research journal. The definitive publisher-authenticated version is available online at the Oxford Journals website. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1313077 |
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