在7M尿素中通过圆盘聚丙烯酰胺凝胶电泳鉴定人酒精脱氢酶同工酶。

Identification of human alcohol dehydrogenase isozymes by disc polyacrylamide gel electrophoresis in 7 M urea.

作者信息

Keung W M, Ditlow C C, Vallee B L

出版信息

Anal Biochem. 1985 Nov 15;151(1):92-6. doi: 10.1016/0003-2697(85)90057-0.

Abstract

Polyacrylamide gel electrophoresis in the presence of 7 M urea provides a simple, reproducible method for the identification of cathodic alcohol dehydrogenase (ADH) isozymes. Treatment of native ADH dimers with 7 M urea and 1 mM dithiothreitol results in a complete dissociation of the 40,000 Mr subunits. Electrophoresis of urea-dissociated ADH isozymes yields a single protein band for homodimers and two bands of equal intensity for heterodimers. The ADH subunits pi, alpha, gamma 2, gamma 1, and beta exhibit electrophoretic mobilities of 0.71, 0.79, 0.88, 0.95, and 1.0, respectively. Thus, the identity of any cathodic ADH isozyme can be determined from the electrophoretic mobilities of its component subunits.

摘要

在7M尿素存在下进行聚丙烯酰胺凝胶电泳，为鉴定阴极醇脱氢酶（ADH）同工酶提供了一种简单、可重复的方法。用7M尿素和1mM二硫苏糖醇处理天然ADH二聚体，会导致40,000 Mr亚基完全解离。尿素解离的ADH同工酶电泳，同型二聚体产生一条单一蛋白带，异型二聚体产生两条强度相等的带。ADH亚基pi、α、γ2、γ1和β的电泳迁移率分别为0.71、0.79、0.88、0.95和1.0。因此，任何阴极ADH同工酶的身份都可以通过其组成亚基的电泳迁移率来确定。