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Interaction of EcoP1 modification methylase with S-adenosyl-L-methionine: a UV-crosslinking study

Krishnamurthy, V and Rao, DN (1994) Interaction of EcoP1 modification methylase with S-adenosyl-L-methionine: a UV-crosslinking study. In: Biochemistry & molecular biology international, 32 (4). 623 -632 .

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Official URL: http://www.ncbi.nlm.nih.gov/pubmed/8038713


EcoP1 modification methylase was radioactively labeled when incubated with S-adenosyl-L-[methyl-3H]methionine in the presence of ultraviolet light. Crosslinking of the enzyme as detected by electrophoresis on sodium dodecyl sulfate-polyacrylamide gel followed by fluorography and autoradiography, was shown to be specific by a number of criteria. More importantly, EcoP1 modification methylase was also radioactively labeled with S-adenosyl-L-[carboxyl-14C]methionine demonstrating that labeling involved binding of the entire AdoMet molecule rather than methylation of the protein. Further, c2 EcoP1 mutant DNA modification methylases which show negligible or very little methylation activity, correspondingly formed a weak or no adduct upon crosslinking. These results suggest that photolabeling of EcoP1 DNA modification methylase occurs at the AdoMet binding site.

Item Type: Journal Article
Publication: Biochemistry & molecular biology international
Publisher: Academic press aust
Additional Information: Copyright of this article belongs to Academic press aust.
Department/Centre: Division of Biological Sciences > Biochemistry
Date Deposited: 13 Apr 2011 05:04
Last Modified: 13 Apr 2011 05:04
URI: http://eprints.iisc.ac.in/id/eprint/36555

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