PMID: 2500935Jan 1, 1989Paper

Localization of a lysine residue near the site of initiating substrate binding of T7 bacteriophage RNA polymerase

Bioorganicheskaia khimiia
T G MaksimoaV P Kumarev

Abstract

A highly selective affinity label was introduced into the T7 phage RNA polymerase by means of GMP ortho-formylphenyl ester and [alpha-32P]UTP nearby the enzyme's active site, which was located using limited cleavage technique. Hydroxylamine, bromine, N-chlorosuccinimide, and cyanogen bromide were employed as the reagents. Analysis of gel-electrophoretic patterns of the cleavage products led to a conclusion that Lys631 is the target of labelling. The region nearby this residue has a high degree of sequence homology with regions of RNA polymerases from T3 and SP6 phages and yeast mitochondria.

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