Regulation of Inwardly Rectifying K^+ Channel in Cultured Opossum Proximal Tubule Cells by Protein Phosphatases 1 and 2A

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Author(s)

Abstract

The inwardly rectifying ATP-regulated K<sup>+</sup> channel with an inward conductance of about 90 pS in the surface membrane of cultured opossum kidney proximal tubule (OKP) cells is activated at least in part by protein kinase A (PKA). In this study, we examined the effects of protein serine/threonine phosphatase types 1 (PP-1) and 2A (PP-2A) on activity of the K<sup>+</sup> channel using the patch-clamp technique. In cell-attached patches, channel activity was enhanced by the application of okadaic acid (OA, 1 μM), a membrane-permeable inhibitor of PP-1 and PP-2A, to the bath solution. This enhancement was abolished by the pretreatment of cells with KT5720 (200 nM), a specific inhibitor of PKA. In inside-out patches, channel activity which could be maintained in the presence of ATP (3 mM) in the bath solution was also increased by the addition of OA (1 μM), and the OA-induced increase in channel activity was partially prevented in the presence of KT5720 (200 nM). Direct application of either PP-1 (1 U/ml) or PP-2A (1 U/ml) to the cytoplasmic surface of the patch membrane inhibited channel activity maintained by ATP (3 mM) in inside-out patches. Moreover, channel activity stimulated by PKA (20 nM) in the presence of ATP (3 mM) was also inhibited by the application of either PP-1 (1 U/ml) or PP-2A (1 U/ml). These results indicate that the OA-sensitive protein phosphatase is involved in the regulation of channel activity, and suggest that both PP-1 and PP-2A are candidates responsible for the inhibition of channel activity through dephosphorylation of the PKA-mediated protein phosphorylation.<br>

Journal

  • The Japanese Journal of Physiology

    The Japanese Journal of Physiology 50(2), 249-256, 2000-04

    THE PHYSIOLOGICAL SOCIETY OF JAPAN

References:  27

Cited by:  1

Codes

  • NII Article ID (NAID)
    10008291095
  • NII NACSIS-CAT ID (NCID)
    AA00691224
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    0021521X
  • NDL Article ID
    5413533
  • NDL Source Classification
    ZS8(科学技術--医学--解剖学・生理学・生化学)
  • NDL Call No.
    Z53-D40
  • Data Source
    CJP  CJPref  NDL  J-STAGE 
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