一种用于检测参与十字花科植物硫代葡萄糖苷生物合成中糖基化和硫酸化步骤的酶的放射分析方法。

A radioassay of enzymes catalyzing the glucosylation and sulfation steps of glucosinolate biosynthesis in Brassica species.

作者信息

Jain J C, Reed D W, GrootWassink J W, Underhill E W

机构信息

Plant Biotechnology Institute, National Research Council, Saskatoon, Saskatchewan, Canada.

出版信息

Anal Biochem. 1989 Apr;178(1):137-40. doi: 10.1016/0003-2697(89)90369-2.

DOI:10.1016/0003-2697(89)90369-2

PMID:2524977

Abstract

A new method for assaying the enzymes uridine diphosphoglucose (UDPglucose):thiohydroximate glucosyltransferase and 3'-phosphoadenosine-5'-phosphosulfate:desulfoglucosinolate sulfotransferase has been designed. The assay system is based on the separation of nonionic [14C]desulfobenzylglucosinolate from anionic [14C]UDPglucose and anionic [14C]benzylglucosinolate, respectively, by differential adsorption to DEAE-ion-exchange disks. The procedure eliminates elaborate chromatographic techniques. The method was used to measure both enzymes in several Brassica spp. In addition, sulfotransferase activity was monitored during partial purification from seedlings of Brassica napus (cv Westar).

摘要

已设计出一种用于测定尿苷二磷酸葡萄糖（UDP葡萄糖）：硫代肟葡萄糖基转移酶和3'-磷酸腺苷-5'-磷酸硫酸酯：脱硫葡萄糖苷磺基转移酶的新方法。该测定系统基于通过对DEAE离子交换盘的差异吸附分别从阴离子型[14C]UDP葡萄糖和阴离子型[14C]苄基葡萄糖苷中分离出非离子型[14C]脱硫苄基葡萄糖苷。该程序省去了复杂的色谱技术。该方法用于测定几种芸苔属植物中的这两种酶。此外，在从甘蓝型油菜（品种Westar）幼苗中进行部分纯化过程中监测了磺基转移酶活性。