Measurement of glycinamide ribonucleotide synthetase activity in intact human fibroblasts and Chinese hamster ovary cells.
Measurement of glycinamide ribonucleotide synthetase activity in intact human fibroblasts and Chinese hamster ovary cells.
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完整人成纤维细胞和中国仓鼠卵巢细胞中甘氨酰胺核糖核苷酸合成酶活性的测量。
DOI：
10.1159/000469333
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Patterson,D
中科院分区：
文献类型：
--
作者：
Hards,RG;Patterson,D
采用基于吐温80渗透性的完整细胞检测系统，研究了人成纤维细胞和中国仓鼠卵巢细胞中甘氨酰胺核糖核苷酸合成酶的活性。确定了测定该酶的最佳条件：三磷酸腺苷、氯化镁、氯化铵、核糖-5‘-磷酸浓度和pH。用Lineweaver-Burke双倒数曲线法测得中国仓鼠卵巢成纤维细胞和中国仓鼠卵巢细胞的V(Max)分别为5.19nmolGAR/5×105Cells/30min和13.4nmolGAR/5×105Cells/30min。
An intact cell assay system based on Tween 80 permeabilization was used to investigate glycinamide ribonucleotide (GAR) synthetase activity in human fibroblasts and Chinese hamster ovary cells. Optimal conditions for the assay of the enzyme were determined with regards to ATP, MgCl(2), NH(4)C1 and ribose-5'-phosphate concentrations as well as pH. Using the optimal assay conditions, the V(max) values as determined by Lineweaver-Burke double reciprocal plots were found to be 5.19 nmol GAR formed/5 X 105 cells/30 min for the fibroblasts and 13.4 nmol GAR formed/5 X 10^5 cells/30 min for the Chinese hamster ovary cells.