EXTRACELLULAR - INTRACELLULAR AND SUBCELLULAR CONCENTRATION GRADIENTS OF THIOLS

被引:30
|
作者
WARDMAN, P
DENNIS, MF
STRATFORD, MRL
WHITE, J
机构
[1] The Gray Laboratory of the Cancer Research Campaign, Mount Vernon Hospital, Northwood, Middlesex HA6 2JR
关键词
THIOLS; GLUTATHIONE; WR-1065; CYSTEINE; INTRACELLULAR CONCENTRATIONS; DNA; ACRIDINE ORANGE; FLUORESCENT PROBES;
D O I
10.1016/0360-3016(92)90517-L
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Chinese hamster V79 cells in Eagle's minimum essential medium in vitro at room temperature were incubated with the aminothiol, WR-1065, or glutathione (GSH) at extracellular concentrations of approximately 1 mmol dm-3. Average intracellular concentrations of GSH, cysteine, and WR-1065 were measured by high performance liquid chromatography, and the effective reducing environment near DNA probed by staining the cells with acridine orange (AO) and measuring the delayed fluorescence. Exposure to either thiol resulted in a rapid, 10-fold increase in average intracellular cysteine concentrations (to about 1 mmol dm-3). Adding extracellular GSH after prior depletion of GSH by treatment with L-buthionine sulfoximine (BSO) did not restore intracellular GSH, but intracellular cysteine was elevated 10-fold. These results are ascribed to thiol/disulfide exchange with cystine in the medium. WR-1065 slowly concentrated intracellularly to approximately 160% of the extracellular concentration. Chemical conjugation of GSH in cells decreased the reducing environment near DNA, but BSO treatment altered the uptake of AO. The electrostatic attraction of WR-1065 toward isolated DNA was markedly affected by ionic strength.
引用
收藏
页码:751 / 754
页数:4
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