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:1979
卷号:76
期号:8
页码:3622-3626
DOI:10.1073/pnas.76.8.3622
语种:English
出版社:The National Academy of Sciences of the United States of America
摘要:Freshly isolated sarcoplasmic reticulum vesicles contain 0.05 mol of tightly bound ADP and 0.03 mol of tightly bound ATP per mol of Ca2+, Mg2+-ATPase (ATP phosphohydrolase, EC 3.6.1.3 ). These values were increased to 0.1-0.2 mol ADP and 0.2-0.3 mol ATP per mol of ATPase after incubation of vesicles in the presence of MgATP and Ca2+ at 25 degrees C and pH 7.0. Half-maximal enrichment of tightly bound nucleotides was obtained with 2.5 mM ATP and 0.32 microM free Ca2+. Uncoupling of calcium transport from ATPase activity by mild acidic conditions or with ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid at pH 7.0 decreased the ability of the membranes to be enriched with tightly bound nucleotides and also decreased the content of tightly bound nucleotides of previously enriched membranes. Tightly bound [3H]nucleotides could only be partially displaced by reincubation under enrichment conditions. Tightly bound nucleotides are associated with energized calcium translocation but do not appear to be directly involved in the catalytic cycle.