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Am J Physiol Renal Physiol 283: F784-F791, 2002. First published June 4, 2002; doi:10.1152/ajprenal.00023.2002
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Vol. 283, Issue 4, F784-F791, October 2002

Protein kinase G activates inwardly rectifying K+ channel in cultured human proximal tubule cells

Kazuyoshi Nakamura, Junko Hirano, Shun-Ichi Itazawa, and Manabu Kubokawa

Department of Physiology II, Iwate Medical University School of Medicine, Morioka, 020-8505 Japan

An ATP-regulated inwardly rectifying K+ channel, whose activity is enhanced by PKA, is present in the plasma membrane of cultured human proximal tubule cells. In this study, we investigated the effects of PKG on this K+ channel, using the patch-clamp technique. In cell-attached patches, bath application of a membrane-permeant cGMP analog, 8-bromoguanosine 3',5'-monophosphate (8-BrcGMP; 100 µM), stimulated channel activity, whereas application of a PKG-specific inhibitor, KT-5823 (1 µM), reduced the activity. Channel activation induced by 8-BrcGMP was observed even in the presence of a PKA-specific inhibitor, KT-5720 (500 nM), which was abolished by KT-5823. Direct effects of cGMP and PKG were examined with inside-out patches in the presence of 1 mM MgATP. Although cytoplasmic cGMP (100 µM) alone had little effect on channel activity, subsequent addition of PKG (500 U/ml) enhanced it. Furthermore, bath application of atrial natriuretic peptide (ANP; 20 nM) in cell-attached patches stimulated channel activity, which was blocked by KT-5823. In conclusion, cGMP/PKG-dependent processes participate in activating the ATP-regulated K+ channel and producing the stimulatory effect of ANP on channel activity.

patch-clamp; human kidney; guanosine 3',5'-cyclic monophosphate; atrial natriuretic peptide; KT-5823


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