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Highly selective affinity labelling of RNA polymerase B (II) from wheat germ

Authors
Journal
FEBS Letters
0014-5793
Publisher
Wiley Blackwell (John Wiley & Sons)
Publication Date
Volume
200
Issue
2
Identifiers
DOI: 10.1016/0014-5793(86)81154-1
Keywords
  • Rna Polymerase B (Ii)(Wheat Germ)Affinity Labeling4-[N-(β-Hydroxyethyl)-N-Methylamino]Benzaldehyded
Disciplines
  • Biology

Abstract

Abstract DNA-dependent RNA polymerase B (II) from wheat germ was modified by incubation with 4-[ N-(β-hydroxyethyl)- N-methyl]benzaldehyde esters of AMP, ADP or ATP, followed by reduction with NaBH 4. Reaction of the modified enzyme with [α- 32P]UTP in the presence of various DNA templates led to a highly selective affinity labelling of the subunit with M r 140000 by covalently linked ApU. Labelling was inhibited by 1 μg ml α-amanitin.

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