Measurement of diacylglycerol acyltransferase activity in isolated hepatocytes

被引：4
|
作者：
Geelen, MJH [1 ]
机构：
[1] Univ Utrecht, Fac Vet Med, Dept Biochem & Cell Biol, NL-3508 TD Utrecht, Netherlands
来源：
ANALYTICAL BIOCHEMISTRY | 2003年 / 322卷 / 02期
关键词：
triacylglycerol synthesis; diacylglycerol acyltransferase; rat hepatocytes; digitonin;
D O I：
10.1016/j.ab.2003.08.019
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
An assay procedure for diacylglycerol acyltransferase that allows rapid measurement of the activity of this enzyme in isolated hepatocytes is described. The one-step procedure involves permeabilization of the plasma membrane with digitonin and simultaneous measurement of diacylglycerol acyltransferase activity. Digitonin at a concentration of 64 mug/mg of cellular protein was found to be optimal for exposing microsomal diacylglycerol acyltransferase to the components of the assay. The enzyme assay is linear with time up to 4 min and with protein concentrations in the range 0.25-2.4 mg of cellular protein/assay. It is shown that there is a good correlation of cellular enzyme activity as determined in digitonin-permeabilized hepatocytes with the rate of triacylglycerol synthesis in intact hepatocytes. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码：264 / 268
页数：5
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