Related Experiment Video
Updated: Aug 9, 2026
A Purification and In Vitro Activity Assay for a (p)ppGpp Synthetase from Clostridium difficile
Published on: November 3, 2018
Partial purification and characterization of cyclic adenosine monophosphate dependent protein kinase from
H Kaur1, S Sharma, G K Khuller
1Department of Biochemistry, Postgraduate Institute of Medical Education and Research, Chandigarh.
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.
