来自大豆幼苗的NAD(P)H：醌氧化还原酶的纯化与特性分析

Purification and characterization of an NAD(P)H:quinone oxidoreductase from Glycine max seedlings.

作者信息

Rescigno A, Sollai F, Masala S, Porcu M C, Sanjust E, Rinaldi A C, Curreli N, Grifi D, Rinaldi A

机构信息

Istituto di Chimica Biologica, Università di Cagliari, Italy.

出版信息

Prep Biochem. 1995 Feb-May;25(1-2):57-67. doi: 10.1080/10826069508010107.

DOI:10.1080/10826069508010107

PMID:7603972

Abstract

An NAD(P)H:(quinone acceptor) oxidoreductase (EC 1.6.99.2) was purified from Glycine max seedlings by means of chromatographic procedures. After 1371-fold purification, the enzyme showed a single band in IEF corresponding to an isoelectric point of 6.1. A single band was also found in native-PAGE both by activity staining and Coomassie brilliant blue staining. The molecular mass determined in SDS-PAGE was 21900 Da, while in HPLC gel-filtration it was 61000 Da. The NAD(P)H:quinone oxidoreductase was able to use NADH or NADPH as the electron donor. Among the artificial quinones which are reduced by this enzyme, 6-hydroxydopa- and 6-hydroxydopamine-quinone are of particular interest because of their neurotoxic effects.

摘要

通过色谱法从大豆幼苗中纯化出一种NAD(P)H：(醌受体)氧化还原酶（EC 1.6.99.2）。经过1371倍纯化后，该酶在等电聚焦中呈现出一条对应于6.1等电点的条带。通过活性染色和考马斯亮蓝染色在非变性聚丙烯酰胺凝胶电泳中也发现了一条单一的条带。在十二烷基硫酸钠聚丙烯酰胺凝胶电泳中测定的分子量为21900道尔顿，而在高效液相色谱凝胶过滤中为61000道尔顿。NAD(P)H：醌氧化还原酶能够使用NADH或NADPH作为电子供体。在被该酶还原的人工醌中，6-羟基多巴醌和6-羟基多巴胺醌因其神经毒性作用而特别受关注。