Orlov S N, Pokudin N I, Kotelevtsev Iu V
Biokhimiia. 1987 Aug;52(8):1373-86.
A rise in intracellular Ca2+(Ca2+in) concentration from 1 to 100 microM is accompanied by a 100-fold increase of erythrocyte membrane permeability for k+ (opening of k+-channels) as well as by membrane hyperpolarization. Both effects are partly inhibited by trifluoroperazine and completely by calmidozolium (R24571). The Ca2+-dependencies of erythrocyte permeability for K+ and of Ca2+ binding to calmodulin are in good correlation. Within the same range of Ca2+in concentrations, i.e. 1-100 microM the activity of Na+-pump decreases by 90% despite the presence of trifluoroperazine and R24571. The permeability of erythrocytes for o-phosphate anions diminishes 15-fold after addition of the anionic exchanger SITS inhibitor. The SITS-inhibited component decreases 9-10 times with a rise in Ca2+in from 10 and 100 microM. In the presence of trifluoroperazine and R24571 the sensitivity of the anionic exchanger towards Ca2+ shows a 2-3 increase. The increase in Ca2+in up to 100 microM is concomitant with the activation of 32Pi incorporation into band 4.1 protein. The effect of Ca2+in on the phosphorylation of this protein is inhibited by calmodulin inhibitors. Addition of protein kinase C activator (4 beta-phorbol-12 beta-myristate-13-acetate) also leads to the increased incorporation of 32P into band 4.1 protein, whereas protein kinase A activator (dibutyryl-cAMP) causes 32P incorporation into bands 4.1 and 5 proteins. No effect of protein kinase activators on the activity of Na+-pump as well as on the permeability of erythrocyte membranes for K+ and anions was revealed. The data obtained point to the differences in the mechanisms of Ca2+in involvement in the regulation of the above ion transport systems. Presumably, none of the mechanisms is coupled with modification of the level of cytoskeleton protein phosphorylation. The effect of Ca2+ is mediated by the Ca2+ interaction with calmodulin only in the case of K+-channels.
细胞内钙离子(Ca2+in)浓度从1微摩尔升至100微摩尔,会伴随红细胞膜对钾离子的通透性增加100倍(钾离子通道开放)以及膜超极化。这两种效应部分被三氟拉嗪抑制,而被卡米多唑(R24571)完全抑制。红细胞对钾离子的通透性以及钙离子与钙调蛋白结合的钙离子依赖性具有良好的相关性。在相同的Ca2+in浓度范围内,即1 - 100微摩尔,尽管存在三氟拉嗪和R24571,钠泵活性仍降低90%。加入阴离子交换剂SITS抑制剂后,红细胞对邻磷酸根阴离子的通透性降低15倍。随着Ca2+in从10微摩尔升至100微摩尔,SITS抑制的成分降低9 - 10倍。在存在三氟拉嗪和R24571的情况下,阴离子交换剂对钙离子的敏感性增加2 - 3倍。Ca2+in增加至100微摩尔会伴随32P掺入带4.1蛋白的激活。Ca2+in对该蛋白磷酸化的作用被钙调蛋白抑制剂抑制。加入蛋白激酶C激活剂(4β - 佛波醇 - 12β - 肉豆蔻酸 - 13 - 乙酸酯)也会导致32P掺入带4.1蛋白增加,而蛋白激酶A激活剂(二丁酰 - cAMP)会使32P掺入带4.1和5蛋白。未发现蛋白激酶激活剂对钠泵活性以及红细胞膜对钾离子和阴离子的通透性有影响。所获得的数据表明Ca2+in参与上述离子转运系统调节的机制存在差异。推测,这些机制均未与细胞骨架蛋白磷酸化水平的改变相关联。仅在钾离子通道的情况下,钙离子的作用是通过钙离子与钙调蛋白的相互作用介导的。