Yamamoto K, Yabu H
Nihon Heikatsukin Gakkai Zasshi. 1980 Jun;16(2):109-21.
Ca2+ binding and ATP dependent Ca2+ uptake by plasma membrane fraction were studied and the effect of treating plasma membrane fraction with trypsin, phospholipase c and neuraminidase on both activities was observed. 1. Plasma membrane fraction possessed the ability to bind Ca2+. The Scatchard plot of Ca2+ binding showed that plasma membrane had at least two types of Ca2+ binding sites of high and low affinity. 2. The amount of Ca2+ uptake by plasma membrane fraction was 18.51 +/- 1.31 nmoles Ca2+/mg protein/30 min at 10(-5)M Ca2+. The Ca2+ concentration for half maximal activation in Ca2+ uptake was 3.7 x 10(-7)M. 3. The ATP dependent Ca2+ uptake by the microsome isolated from guinea pig intestine was clearly stimulated by 4 mM K2-oxalate whereas that of plasma membrane fraction was not. 4. Trypsin and phospholipase c treatment led to a 40--88% reduction in the ATP dependent Ca2+ uptake. The Ca2+ concentration for half maximal activation Ca2+ uptake shifted to a high concentration. On the other hand, neuraminidase treatment resulted in a 43--93% increase in ATP dependent Ca2+ uptake. But, the Ca2+ concentration for half maximal activation in Ca2+ uptake was not shifted by neuraminidase treatment. The results show that plasma membrane plays an important role in regulating the cytoplasmic Ca2+ concentration. The results also suggest that a glycoprotein linked with sialic acid is involved in the flux of Ca2+ across the plasma membrane.
研究了质膜组分对Ca2+的结合以及ATP依赖的Ca2+摄取情况,并观察了用胰蛋白酶、磷脂酶c和神经氨酸酶处理质膜组分对这两种活性的影响。1. 质膜组分具有结合Ca2+的能力。Ca2+结合的Scatchard图表明,质膜至少有两种高亲和力和低亲和力的Ca2+结合位点。2. 在10(-5)M Ca2+浓度下,质膜组分摄取Ca2+的量为18.51±1.31 nmol Ca2+/mg蛋白质/30分钟。Ca2+摄取半最大激活浓度为3.7×10(-7)M。3. 从豚鼠肠道分离的微粒体中ATP依赖的Ca2+摄取明显受到4 mM草酸钾的刺激,而质膜组分则不受影响。4. 胰蛋白酶和磷脂酶c处理导致ATP依赖的Ca2+摄取减少40%-88%。Ca2+摄取半最大激活的Ca2+浓度移向高浓度。另一方面,神经氨酸酶处理导致ATP依赖的Ca2+摄取增加43%-93%。但是,神经氨酸酶处理并未使Ca2+摄取半最大激活的Ca2+浓度发生改变。结果表明,质膜在调节细胞质Ca2+浓度方面起重要作用。结果还表明,与唾液酸相连的糖蛋白参与Ca2+跨质膜的通量。