Prishchepa L A, Burdyga F V, Kosterin S A
Biokhimiia. 1996 Jul;61(7):1250-6.
The effects of Ca(2+)-precipitating anions (oxalate and phosphate) and effective inhibitors of endo/sarcoplasmic reticulum calcium pump (thapsigargin and cyclopiazonic acid) on azide-insensitive (5 mM) Mg2+,ATP-dependent Ca2+ accumulation in microsomes of ureter smooth muscle cells were studied. Oxalate (0-20 mM) and phosphate (0-60 mM) stimulate Mg2+,ATP-dependent Ca2+ accumulation. Thapsigargin and cyclopiazonic acid at 100 nM and 20 microM, respectively, completely inhibited (i.e., down to the level in the absence of oxalate) Ca2+ accumulation activated by 10 nM oxalate. These inhibitors only partially inhibited Ca2+ accumulation activated by 40 mM phosphate. Mg2+,ATP-dependent Ca2+ accumulation in microsomes, which is inhibited by thapsigargin and cyclopiazonic acid and activated by oxalate or phosphate, can result from functioning of calcium pump in endoplasmic reticulum of ureter myocytes. The inhibition constant, Ki, was calculated by the method of Hill and it was 0.3 nM and 0.2 microM for thapsigargin or cyclopiazonic acid, respectively. Mg2+,ATP-dependent Ca2+ accumulation in microsomes, which is insensitive to thapsigargin or cyclopiazonic acid and activated by phosphate, can result from functioning of calcium pump in plasma membranes of ureter myocytes.
研究了钙离子沉淀阴离子(草酸盐和磷酸盐)以及内质网/肌浆网钙泵的有效抑制剂(毒胡萝卜素和环匹阿尼酸)对输尿管平滑肌细胞微粒体中叠氮化物不敏感(5 mM)的镁离子、ATP 依赖性钙离子积累的影响。草酸盐(0 - 20 mM)和磷酸盐(0 - 60 mM)刺激镁离子、ATP 依赖性钙离子积累。毒胡萝卜素和环匹阿尼酸分别在 100 nM 和 20 μM 时,完全抑制(即降至无草酸盐时的水平)由 10 nM 草酸盐激活的钙离子积累。这些抑制剂仅部分抑制由 40 mM 磷酸盐激活的钙离子积累。微粒体中镁离子、ATP 依赖性钙离子积累,被毒胡萝卜素和环匹阿尼酸抑制并被草酸盐或磷酸盐激活,可能是输尿管肌细胞内质网中钙泵发挥作用的结果。通过希尔方法计算抑制常数 Ki,毒胡萝卜素或环匹阿尼酸的 Ki 分别为 0.3 nM 和 0.2 μM。微粒体中镁离子、ATP 依赖性钙离子积累,对毒胡萝卜素或环匹阿尼酸不敏感并被磷酸盐激活,可能是输尿管肌细胞质膜中钙泵发挥作用的结果。