Atrial natriuretic peptide-stimulated Ca2+ reabsorption in rabbit kidney requires membrane-targeted, cGMP-dependent protein kinase type II

被引:43
作者
Hoenderop, JGJ
Vaandrager, AB
Dijkink, L
Smolenski, A
Gambaryan, S
Lohmann, SM
de Jonge, HR
Willems, PHGM
Bindels, RJM
机构
[1] Univ Nijmegen, Inst Cellular Signaling, Dept Biochem, NL-6500 HB Nijmegen, Netherlands
[2] Univ Nijmegen, Inst Cellular Signaling, Dept Cell Physiol, NL-6500 HB Nijmegen, Netherlands
[3] Erasmus Univ, Dept Biochem, Cardiovasc Res Inst, COEUR, NL-3000 DR Rotterdam, Netherlands
[4] Univ Wurzburg, Med Clin, Lab Clin Biochem, D-97080 Wurzburg, Germany
关键词
cortical collecting duct; connecting tubule; adenovirus; sodium transport; ENaC;
D O I
10.1073/pnas.96.11.6084
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Atrial natriuretic peptide (ANP) and nitric oxide (NO) are key regulators of ion and water transport in the kidney. Here, we report that these cGMP-elevating hormones stimulate Ca2+ reabsorption via a novel mechanism specifically involving type II cGMP-dependent protein kinase (cGK II). ANP and the NO donor, sodium nitroprusside (SNP), markedly increased Ca2+ uptake in freshly immunodissected rabbit connecting tubules (CNT) and cortical collecting ducts (CCD). Although readily increasing cGMP, ANP and SNP did not affect Ca2+ and Na+ reabsorption in primary cultures of these segments. Immunoblot analysis demonstrated that cGK II, and not cGK I, was present in freshly isolated CNT and CCD but underwent a complete down-regulation during the primary cell culture. However, upon adenoviral reexpression of cGK II in primary cultures, ANP, SNP, and 8-Br-cGMP readily increased Ca2+ reabsorption. In contrast, no cGMP-dependent effect on electrogenic Na+ transport was observed. The membrane localization of cGK II proved to be crucial for its action, because a nonmyristoylated cGK II mutant that was shown to be localized in the cytosol failed to mediate ANP-stimulated Ca2+ transport. The Ca2+-regulatory function of cGK II appeared isotype-specific because no cGMP-mediated increase in Ca2+ transport was observed after expression of the cytosolic cGK IP or a membrane-bound cGK II/I beta chimer. These results demonstrate that ANP- and NO-stimulated Ca2+ reabsorption requires membrane-targeted cGK II.
引用
收藏
页码:6084 / 6089
页数:6
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