Eilam Y, Othman M, Halachmi D
Department of Bacteriology, Hebrew University, Hadassah Medical School, Jerusalem, Israel.
J Gen Microbiol. 1990 Dec;136(12):2537-43. doi: 10.1099/00221287-136-12-2537.
Influx of 45Ca2+ into Saccharomyces cerevisiae was measured under experimental conditions which enabled measurements of initial rate of transport across the plasma membrane, without interference by the vacuolar Ca2+ transport system. Addition of glucose or glycerol to the cells, after pre-incubation in glucose-free medium for 5 min, caused a rapid, transient increase in 45Ca2+ influx, reaching a peak at 3-5 min after addition of substrate. Ethanol, or glycerol added with antimycin A, had no effect on 45Ca2+ influx. We have shown previously that this increase is not mediated by an effect of the substrates on intracellular ATP levels. Changes in membrane potential accounted for only a part of the glucose-stimulated 45Ca2+ influx. The roles of intracellular acidification and changes in cellular cAMP in mediating the effects of glucose on 45Ca2+ influx were examined. After a short preincubation in glucose-free medium addition of glucose caused a decrease in the intracellular pH, [pH]i, which reached a minimum value after 3 min. A transient increase in the cellular cAMP level was also observed. Addition of glycerol also caused intracellular acidification, but ethanol or glycerol added with antimycin A had no effect on [pH]i. Artificial intracellular acidification induced by exposure to isobutyric acid or to CCCP caused a transient rise in Ca2+ influx but the extent of the increase was smaller than that caused by glucose, and the time-course was different. We conclude that intracellular acidification may be responsible for part of the glucose stimulation of Ca2+ influx.(ABSTRACT TRUNCATED AT 250 WORDS)
在能够测量穿过质膜的初始运输速率且不受液泡钙运输系统干扰的实验条件下,测定了45Ca2+流入酿酒酵母的情况。在无葡萄糖培养基中预孵育5分钟后,向细胞中添加葡萄糖或甘油,会导致45Ca2+流入迅速、短暂增加,在添加底物后3 - 5分钟达到峰值。乙醇或与抗霉素A一起添加的甘油对45Ca2+流入没有影响。我们之前已经表明,这种增加不是由底物对细胞内ATP水平的影响介导的。膜电位的变化仅占葡萄糖刺激的45Ca2+流入的一部分。研究了细胞内酸化和细胞cAMP变化在介导葡萄糖对45Ca2+流入影响中的作用。在无葡萄糖培养基中短暂预孵育后添加葡萄糖会导致细胞内pH值([pH]i)下降,3分钟后达到最小值。还观察到细胞cAMP水平短暂升高。添加甘油也会导致细胞内酸化,但乙醇或与抗霉素A一起添加的甘油对[pH]i没有影响。暴露于异丁酸或CCCP诱导的人工细胞内酸化会导致Ca2+流入短暂上升,但增加的程度小于葡萄糖引起的,且时间进程不同。我们得出结论,细胞内酸化可能是葡萄糖刺激Ca2+流入的部分原因。(摘要截短至250字)