(Pro)renin receptor is required for prorenin-dependent and -independent regulation of vacuolar H+-ATPase activity in MDCK.C11 collecting duct cells
(Pro)renin receptor is required for prorenin-dependent and -independent regulation of vacuolar H+-ATPase activity in MDCK.C11 collecting duct cells
复制标题
DOI：
10.1152/ajprenal.00037.2013
复制
发表时间：
2013-08-01
影响因子：
4.2
通讯作者：
Meima, Marcel E.
中科院分区：
文献类型：
--
作者：
Lu, Xifeng;Garrelds, Ingrid M.;Meima, Marcel E.
Prorenin与Prorenin受体结合[(P)RR]导致Prorenin的非蛋白水解激活，但也直接（即独立于血管紧张素的产生）激活信号转导级联，从而导致促纤维化因子的上调。(P)RR是液泡型H+- atp酶（v - atp酶）的辅助蛋白，是v - atp酶完整性所必需的。此外，在收集管细胞中，催乳素诱导的Erk活化依赖于v - atp酶活性。然而，与(P)RR结合的prorenin是否直接调节V-ATPase活性尚不清楚。本实验研究了prorenin对Madin-Darby犬肾克隆11 （MDCK.C11）细胞质膜v - atp酶活性的影响。在低纳摩尔浓度下，Prorenin增加v - atp酶的活性，v - atp酶抑制剂巴菲霉素A1，而血管紧张素II型1和2受体阻滞剂伊贝沙坦和PD-123319不能阻止这种情况。增加的v - atp酶活性，但不是基础，被(P)RR的小干扰RNA耗尽所消除。出乎意料的是，假定的肽(P)RR阻滞剂柄区肽也以(P)RR依赖的方式增加v - atp酶活性。最后，[Arg(8)]-抗利尿激素刺激的V-ATPase活性和cAMP的产生也被(P)RR消耗所消除。我们的结果表明，在MDCK。在C11细胞中，(P)RR对V-ATPase活性的依赖性和非依赖性调节是必需的。
Prorenin binding to the prorenin receptor [(P)RR] results in nonproteolytic activation of prorenin but also directly (i.e., independent of angiotensin generation) activates signal transduction cascades that can lead to the upregulation of profibrotic factors. The (P)RR is an accessory protein of vacuolar-type H+-ATPase (V-ATPase) and is required for V-ATPase integrity. In addition, in collecting duct cells, prorenin-induced activation of Erk depends on V-ATPase activity. However, whether prorenin binding to the (P)RR directly regulates V-ATPase activity is as yet unknown. Here, we studied the effect of prorenin on plasma membrane V-ATPase activity in Madin-Darby canine kidney clone 11 (MDCK.C11) cells, which resemble intercalated cells of the collecting duct. Prorenin increased V-ATPase activity at low nanomolar concentrations, and the V-ATPase inhibitor bafilomycin A1, but not the angiotensin II type 1 and 2 receptor blockers irbesartan and PD-123319, prevented this. Increased, but not basal, V-ATPase activity was abolished by small interfering RNA depletion of the (P)RR. Unexpectedly, the putative peptidic (P)RR blocker handle region peptide also increased V-ATPase activity in a (P)RR-dependent manner. Finally, [Arg(8)]-vasopressin-stimulated V-ATPase activity and cAMP production were also abolished by (P)RR depletion. Our results show that in MDCK.C11 cells, the (P)RR is required for prorenin-dependent and -independent regulation of V-ATPase activity.