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

[摘要]

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 NaBH₄. 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 α-amanitin.

[学科分类] 生物化学/生物物理

[关键词] RNA polymerase B (II) (Wheat germ) Affinity labeling 4-[N-(β-Hydroxyethyl)-N-methylamino]benzaldehyde Dinucleotide synthesis