Open AccessPurification and characterization of a tRNA methylase from Salmonella typhimurium
Author(s) -
William T. Pope,
Robert H. Reeves
Publication year - 1978
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.136.1.191-200.1978
Subject(s) - biology , enzyme , sephadex , biochemistry , transfer rna , gel electrophoresis , salmonella , polyacrylamide gel electrophoresis , sodium dodecyl sulfate , escherichia coli , enterobacteriaceae , column chromatography , bacteria , chromatography , microbiology and biotechnology , rna , chemistry , gene , genetics
A tRNA methylase, in which supK strains of Salmonella typhimurium are deficient, was purified from strain LT2 and characterized. Column chromatography of protein extracts from wild-type cells on phosphocellulose, diethylaminoethyl-Sephadex A-50, and hydroxlapatite resulted in an enzyme that was estimated to be about 50% pure. tRNA from S. typhimurium which had been incubated at pH 9.0 served as a substrate for this methylase. The enzyme has a molecular weight of about 50,000 as estimated by gel chromatography and by electrophoresis on sodium dodecyl sulfate-polyacrylamide gels. The optimal assay conditions, as well as the kinetics and stability of the enzyme, were studied. As with other tRNA-methylating enzymes, S-adenosylhomocysteine is a potent inhibitor.
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