Grover A K, Samson S E, Berenski C J, Jung C Y
Life Sci. 1985 Dec 9;37(23):2193-8. doi: 10.1016/0024-3205(85)90571-5.
The target sizes of the oxalate-independent Ca uptake by the plasma membrane enriched fraction F2, and the oxalate-stimulated Ca uptake by a fraction F3 slightly enriched in the endoplasmic reticulum were determined by radiation inactivation. The oxalate-independent Ca uptake was inactivated with a D37 value of 1.96 +/- 0.30 Mrad but the oxalate-stimulated Ca uptake had a D37 value of 0.45 +/- 0.07 Mrad. Thus, in the smooth muscle the oxalate-stimulated Ca uptake appeared to be due to a structure 3 to 6 times larger than was the oxalate-independent Ca uptake. The subcellular site of the ATP-dependent azide insensitive Ca uptake in the smooth muscle has been disputed in the past. It has been suggested to be plasma membrane (PM) by several workers, and endoplasmic reticulum (ER) by others. Recently, however, there has been substantial evidence to support the hypothesis that one Ca uptake system, unaffected by oxalate, resides in the PM and another, stimulated by oxalate, is located in the ER of the smooth muscle. The evidence has been reviewed recently. Here, we show that the two modes of Ca uptake differ in their target sizes as well. To our knowledge, this is the first report on the use of radiation inactivation to distinguish between the two modes of Ca uptake in any tissue.
通过辐射失活法测定了富含质膜的组分F2对草酸盐不依赖的钙摄取的靶标大小,以及内质网轻度富集的组分F3对草酸盐刺激的钙摄取的靶标大小。草酸盐不依赖的钙摄取的D37值为1.96±0.30兆拉德时被失活,但草酸盐刺激的钙摄取的D37值为0.45±0.07兆拉德。因此,在平滑肌中,草酸盐刺激的钙摄取似乎是由于一种比草酸盐不依赖的钙摄取大3至6倍的结构所致。过去,平滑肌中依赖ATP的叠氮化物不敏感的钙摄取的亚细胞位点一直存在争议。一些研究人员认为是质膜(PM),另一些人则认为是内质网(ER)。然而,最近有大量证据支持这样一种假说,即一种不受草酸盐影响的钙摄取系统存在于质膜中,另一种受草酸盐刺激的钙摄取系统位于平滑肌的内质网中。最近对这些证据进行了综述。在这里,我们表明这两种钙摄取模式在其靶标大小上也有所不同。据我们所知,这是关于使用辐射失活法区分任何组织中两种钙摄取模式的首次报道。