小麦胚芽RNA聚合酶B（II）的高度选择性亲和标记

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

作者信息

Grachev M A, Hartmann G R, Maximova T G, Mustaev A A, Schäffner A R, Sieber H, Zaychikov E F

出版信息

FEBS Lett. 1986 May 12;200(2):287-90. doi: 10.1016/0014-5793(86)81154-1.

DOI:10.1016/0014-5793(86)81154-1

Abstract

DNA-dependent RNA polymerase B (II) from wheat germ was modified by incubation with 4-[N-(beta-hydroxyethyl)-N-methyl]benzaldehyde esters of AMP, ADP or ATP, followed by reduction with NaBH4. Reaction of the modified enzyme with [alpha-32P]UTP in the presence of various DNA templates led to a highly selective affinity labelling of the subunit with Mr 140 000 by covalently linked ApU. Labelling was inhibited by 1 microgram/ml alpha-amanitin.

摘要

从小麦胚芽中提取的依赖DNA的RNA聚合酶B（II），通过与AMP、ADP或ATP的4-[N-（β-羟乙基）-N-甲基]苯甲醛酯一起温育，随后用NaBH4还原进行修饰。在各种DNA模板存在的情况下，修饰后的酶与[α-32P]UTP反应，通过共价连接的ApU对分子量为140000的亚基进行高度选择性的亲和标记。1微克/毫升的α-鹅膏蕈碱可抑制标记。

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