A HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY-BASED RADIOMETRIC ASSAY FOR SUCROSE-PHOSPHATE SYNTHASE AND OTHER UDP-GLUCOSE REQUIRING ENZYMES

被引：7

作者：

SALVUCCI, ME ^{[1
]}

CRAFTSBRANDNER, SJ ^{[1
]}

机构：

[1] UNIV KENTUCKY,DEPT AGRON,LEXINGTON,KY 40546

来源：

ANALYTICAL BIOCHEMISTRY | 1991年 / 194卷 / 02期

关键词：

D O I：

10.1016/0003-2697(91)90242-L

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method for product analysis that eliminates a problematic step in the radiometric sucrose-phosphate synthase assay is decribed. The method uses chromatography on a boronate-derivatized high-performance liquid chromatography column to separate the labeled product, [14]sucrose phosphate, from unreacted uridine 5′-diphosphate-[14C]glucose (UDP-Glc). Direct separation of these compounds eliminates the need for treatment of the reaction mixtures with alkaline phosphatase, thereby avoiding the problem of high background caused by contaminating phosphodiesterase activity in alkaline phosphatase preparations. The method presented in this paper can be applied to many UDP-Glc requiring enzymes; here we show its use for determining the activities of sucrose-phosphate synthase, sucrose synthase, and uridine diphosphate-glucose pyrophosphorylase in plant extracts. © 1991.

引用

页码：365 / 368

页数：4

共 15 条

[1] KINETIC CHARACTERIZATION OF SPINACH LEAF SUCROSE-PHOSPHATE SYNTHASE [J].