z-logo
open-access-imgOpen Access
Purification of theE.coliogt gene product to homogeneity and its rate of action onO6-methylguanine,O6-ethyiguanine andO4-methylthymine in dodecadeoxyribnucleotides
Author(s) -
Mark C. Wilkinson,
Philip M. Potter,
Lynn Cawkwell,
Panagiotis Georgiadis,
Dilip D. Patel,
Peter Swann,
Geoffrey P. Margison
Publication year - 1989
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/17.21.8475
Subject(s) - biology , microbiology and biotechnology , escherichia coli , gene , genetics
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 (approximately 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 greater than O6-EtG greater than O4-MeT, however, the ogt protein was found to repair O6-MeG, O6-EtG and O4-Met, 1.1, 173 and 84 times, respectively, faster than the ada protein.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.
Having issues? You can contact us here