title: Partial purification and characterization of cyclic adenosine monophosphate dependent protein kinase from mycobacterium smegmatis.
abstract: Adenosine 3' 5'-monophosphate (cyclic AMP) - dependent protein kinase was purified about 42 fold from the M. smegmatis by ammonium sulphate fractionation followed by DEAE-cellulose and phosphocellulose column chromatography. SDS-PAGE revealed two prominent bands, with molecular masses of 55 KDa and 58 KDa. The enzyme preferentially utilized phosvitin and Histones as exogenous phosphate acceptor. Mg2+ ions were essential for enzyme activity, other metal ions like Ca2+, Zn2+, Co2+ and Mn2+, could not substitute for Mg2+. Inhibition of enzyme activity by thiol reagents, 5-5'-dithio bis (2-nitrobenzoic acid) and N-ethylmaleimide suggest that the cystein residues in the protein kinase might be located at or near the active site of the enzyme.
authors.affiliation: Department of Biochemistry, Postgraduate Institute of Medical Education and Research, Chandigarh.
authors.forename: H
authors.initials: H
authors.lastname: Kaur
authors.forename: S
authors.initials: S
authors.lastname: Sharma
authors.forename: G K
authors.initials: GK
authors.lastname: Khuller
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002852:Chromatography, Ion Exchange; D000242:Cyclic AMP / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism*; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005033:Ethylmaleimide / Q000494:pharmacology; D006657:Histones / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009161:Mycobacterium / Q000201:enzymology*; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation; D010774:Phosvitin / Q000378:metabolism; D013379:Substrate Specificity; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013696:Temperature
chemical_list: D006657:Histones; D008670:Metals; D010455:Peptides; D013439:Sulfhydryl Reagents; D010774:Phosvitin; D004228:Dithionitrobenzoic Acid; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D005033:Ethylmaleimide