[Localization of a lysine residue near the site of initiating substrate binding of T7 bacteriophage RNA polymerase].
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.
Maksimoa
T G
TG
Mustaev
A A
AA
Zaĭchikov
E F
EF
Baranova
L V
LV
Kumarev
V P
VP
D000345:Affinity Labels; D001665:Binding Sites; D001966:Bromine; D003488:Cyanogen Bromide; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D006898:Hydroxylamines; D008239:Lysine / Q000032:analysis*; D008969:Molecular Sequence Data; D012689:Sequence Homology, Nucleic Acid; D013388:Succinimides; D013604:T-Phages / Q000201:enzymology*
D000345:Affinity Labels; D006898:Hydroxylamines; D013388:Succinimides; C015752:N-chlorosuccinimide; D012321:DNA-Directed RNA Polymerases; D008239:Lysine; D003488:Cyanogen Bromide; D001966:Bromine
Lokaliatsiia ostatka lizina v oblasti sviazyvaniia initsiruiushchego substrata RNK-polimerazy bakteriofaga T7.