Sze H, Solomon A K
Biochim Biophys Acta. 1979 Jun 13;554(1):180-94. doi: 10.1016/0005-2736(79)90017-8.
We have characterized the asymmetric effect of Ca2+ on passive K+ permeability in erythrocyte membranes, using inside out and right-side out vesicles. Ca2+, but not Mg2+, can induce an increase in K+ uptake in inside out vesicles. The half-maximal concentration of Ca2+ required to induce the K+ uptake is 0.2 mM, and the permeability increase is not specific for K+. Thus, the Ca2+- induced permeation process in inside out vesicles is changed from that in the energy-depleted intact cell which requires only micromolar concentrations of Ca2+ and is specific for K+. Removal of spectrin had no effect on the vesicle permeability increase due to Ca2+. Studies with N-ethylmaleimide show that the vesicle channel openings is mediated by a protein and passage is controlled by sulfhydryl groups; furthermore, the Ca2+-induced vesicle pathway is distinct from the normal channel for passive K+ leak in the absence of Ca2+. The protein is sensitive to its phospholipid environment since removal of easily accessible phospholipid head groups on the cytoplasmic face of the vesicles inhibits the Ca2+ -stimulated channel opening.
我们使用内翻式和外翻式囊泡,表征了Ca2+对红细胞膜被动K+通透性的不对称效应。Ca2+而非Mg2+可诱导内翻式囊泡中K+摄取增加。诱导K+摄取所需的Ca2+半数最大浓度为0.2 mM,且通透性增加并非K+所特有。因此,内翻式囊泡中Ca2+诱导的通透过程与能量耗尽的完整细胞中的情况不同,后者仅需要微摩尔浓度的Ca2+,且对K+具有特异性。去除血影蛋白对Ca2+引起的囊泡通透性增加没有影响。用N-乙基马来酰亚胺进行的研究表明,囊泡通道开放由一种蛋白质介导,且通道通过受巯基控制;此外,Ca2+诱导的囊泡通道与无Ca2+时被动K+泄漏的正常通道不同。该蛋白质对其磷脂环境敏感,因为去除囊泡胞质面上易于接近的磷脂头部基团会抑制Ca2+刺激的通道开放。