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Bioorg Khim ; 16(8): 1145-8, 1990 Aug.
Article in Russian | MEDLINE | ID: mdl-2285430

ABSTRACT

RNA polymerase II from human placenta was affinity labelled in crude preparation using two-step technique, which includes treatment of the enzyme with an aldehyde-containing reactive analogue of ATP, ADP or AMP in the presence of poly[d(A-T)] followed (after borohydride reduction) by the elongation of the attached label with [alpha-32P]UTP. A polypeptide of the molecular mass ca. 140 kDa proved to be the labelling target. No labelling was observed in the absence of poly[d(A-T)] or the reagent or in the presence of alpha-amanitin. All the results suggest the attachment of the affinity reagents to the second-largest subunit of the human RNA polymerase II, which therefore takes part in the initiation substrate's binding.


Subject(s)
Placenta/enzymology , RNA Polymerase II/chemistry , Affinity Labels , Binding Sites , Chromatography, DEAE-Cellulose , Electrophoresis, Polyacrylamide Gel , Female , Humans
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