期刊名称:Proceedings of the National Academy of Sciences
印刷版ISSN:0027-8424
电子版ISSN:1091-6490
出版年度:1974
卷号:71
期号:12
页码:5042-5046
DOI:10.1073/pnas.71.12.5042
语种:English
出版社:The National Academy of Sciences of the United States of America
摘要:Accumulation of 45Ca++ was found to occur in membrane vesicles of E. coli prepared by lysis with a French pressure cell. The uptake occurs by active transport, requiring an energy source. Substrates of the electron transport chain, including D-lactate, reduced phenazine methosulfate, and NADH, stimulated accumulation, but this effect was blocked by the addition of cyanide. ATP could also stimulate accumulation, and this effect was blocked by dicyclohexylcarbodiimide. Uncouplers of oxidative phosphorylation inhibited the accumulation driven by either type of energy source. Accumulation of calcium is rapid, reaching the steady-state plateau within 1 min. Addition of phosphate to the assay buffer results in a prolongation of the reaction, allowing for the time-dependent accumulation of calcium for as long as 30 min. Vesicles prepared by lysis with a French pressure cell exhibit almost no ability to accumulate proline, while vesicles prepared by the method of Kaback transport proline but exhibit little energy-dependent transport of calcium. It is suggested that the accumulation of calcium in these vesicles, which are believed to be inverted, reflects a system that in vivo is responsible for the active extrusion of calcium from the cells.
关键词:membrane orientation ; energy coupling ; calcium uptake