Smith H G, Capalbo P M
Biochemistry. 1985 Jul 16;24(15):4233-9. doi: 10.1021/bi00336a063.
Increasing the concentration of calcium in the external buffer flowing past isolated, intact bovine retinal rod outer segment disks immobilized in a flow system reduced the rate of radioactive calcium efflux from within the disks in the dark. We interpret these results as extradiskal calcium acting at an inhibitory binding site to block the calcium efflux. A Scatchard analysis of the external calcium dependence of the efflux yields an apparent dissociation constant of 50 microM, which further suggests that the inhibition is mediated by a specific membrane binding site. The observed inhibition of calcium efflux may represent a functional role for the high-affinity calcium binding site which has been identified by others in previous physical studies of the disk membrane. This external calcium inhibited permeability may explain some of the discrepancies in the reported calcium transport properties of disks. Variations in the external calcium concentration may alter the calcium content of isolated disks, thereby indirectly affecting other transport functions including the measured light-induced release of calcium. No evidence was found for either Na/Ca or Ca/Ca exchange processes across the disk membrane. Lanthanum was even more effective than calcium in inhibiting calcium efflux in the dark. Neither lanthanum nor calcium inhibited the light-induced efflux of calcium from disks, which implies either that light and extradiskal calcium regulate separate permeability processes in the disk membrane or that light greatly reduces the affinity of the inhibitory site for calcium and lanthanum.
增加流过固定在流动系统中的分离、完整的牛视网膜杆外段圆盘的外部缓冲液中的钙浓度,会降低黑暗中圆盘内放射性钙的流出速率。我们将这些结果解释为盘外钙作用于一个抑制性结合位点以阻断钙流出。对流出的外部钙依赖性进行Scatchard分析得出表观解离常数为50微摩尔,这进一步表明抑制作用是由一个特定的膜结合位点介导的。观察到的钙流出抑制可能代表了高亲和力钙结合位点的功能作用,该位点已在之前对圆盘膜的物理研究中被其他人鉴定出来。这种外部钙抑制的通透性可能解释了报道的圆盘钙转运特性中的一些差异。外部钙浓度的变化可能会改变分离圆盘的钙含量,从而间接影响其他转运功能,包括测量到的光诱导钙释放。未发现跨圆盘膜存在钠/钙或钙/钙交换过程的证据。镧在黑暗中比钙更有效地抑制钙流出。镧和钙都不抑制光诱导的圆盘钙流出,这意味着要么光和盘外钙调节圆盘膜中不同的通透性过程,要么光大大降低了抑制位点对钙和镧的亲和力。