Morel N, Wibo M, Godfraind T
Biochim Biophys Acta. 1981 Jun 9;644(1):82-8. doi: 10.1016/0005-2736(81)90061-4.
An ATP-driven Ca2+-transport system has been characterized in a microsomal fraction from rat aorta. Calmodulin enhanced 2.5-fold 45Ca accumulation by EGTA-treated microsomes incubated with 10 microM Ca2+ (in the absence of oxalate) by increasing markedly the apparent affinity of the transport system for Ca2+. The ionophore A23187 induced a rapid release of the sequestered 45Ca. The vesicles that took up 45Ca were distributed like plasmalemmal marker enzymes when the microsomal fraction was subfractionated by density gradient centrifugation. In particular, these vesicles were markedly shifted towards higher equilibrium densities after addition to the microsomes of 0.2 mg digitonin/mg protein before isopycnic centrifugation. We conclude that the calmodulin-stimulated Ca2+ pump associated with the microsomal fraction is located in plasmalemmal elements.
已在大鼠主动脉微粒体组分中对一种由ATP驱动的Ca2+转运系统进行了表征。钙调蛋白通过显著提高转运系统对Ca2+的表观亲和力,使经EGTA处理并与10微摩尔Ca2+(无草酸盐存在)孵育的微粒体的45Ca积累增加了2.5倍。离子载体A23187诱导了被隔离的45Ca的快速释放。当通过密度梯度离心对微粒体组分进行亚分级分离时,摄取45Ca的囊泡的分布与质膜标记酶相似。特别是,在等密度离心前向微粒体中加入0.2毫克洋地黄皂苷/毫克蛋白质后,这些囊泡明显向更高的平衡密度移动。我们得出结论,与微粒体组分相关的钙调蛋白刺激的Ca2+泵位于质膜成分中。