ATP receptor regulation of adenylate cyclase and protein kinase C activity in cultured renal LLC-PK1 cells

Robert J Anderson; Ruth Breckon; Bradley S Dixon

doi:10.1172/JCI115191

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ATP receptor regulation of adenylate cyclase and protein kinase C activity in cultured renal LLC-PK1 cells

Journal article

Open access

Peer reviewed

ATP receptor regulation of adenylate cyclase and protein kinase C activity in cultured renal LLC-PK1 cells

Robert J Anderson, Ruth Breckon and Bradley S Dixon

The Journal of clinical investigation, Vol.87(5), pp.1732-1738

05/1991

DOI: 10.1172/JCI115191

PMCID: PMC295279

PMID: 1850760

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Abstract

In cultured intact LLC-PK1 renal epithelial cells, a nonhydrolyzable ATP analogue, ATP gamma S, inhibits AVP-stimulated cAMP formation. In LLC-PK1 membranes, several ATP analogues inhibit basal, GTP-, forskolin-, and AVP-stimulated adenylate cyclase activity in a dose-dependent manner. The rank order potency of inhibition by ATP analogues suggests that a P2y type of ATP receptor is involved in this inhibition. The compound ATP gamma S inhibits agonist-stimulated adenylate cyclase activity in solubilized and in isobutylmethylxanthine (IBMX) and quinacrine pretreated membranes, suggesting that ATP gamma S inhibition occurs independent of AVP and A1 adenosine receptors and of phospholipase A2 activity. The ATP gamma S inhibition of AVP-stimulated adenylate cyclase activity is not affected by pertussis toxin but is attenuated by GDP beta S, suggesting a possible role for a pertussis toxin insensitive G protein in the inhibition. Exposure of intact LLC-PK cells to ATP gamma S results in a significant increase in protein kinase C activity. However, neither of two protein kinase C inhibitors (staurosporine and H-7) prevents ATP gamma S inhibition of AVP-stimulated adenylate cyclase activity, suggesting that this inhibition occurs by a protein kinase C independent mechanism. These findings suggest the presence of functional P2y purinoceptors coupled to two signal transduction pathways in cultured renal epithelial cells. The effect of P2y purinoceptors to inhibit AVP-stimulated adenylate cyclase activity may be mediated, at least in part, by a pertussis toxin insensitive G protein.

Adenosine Triphosphate - pharmacology

Receptors, Purinergic - physiology

3',5'-Cyclic-AMP Phosphodiesterases - analysis

Adenylyl Cyclases - analysis

GTP-Binding Proteins - physiology

Cyclic AMP - biosynthesis

Kidney - enzymology

Cells, Cultured

Receptors, Purinergic - drug effects

Arginine Vasopressin - pharmacology

Adenosine Triphosphate - analogs & derivatives

Protein Kinase C - analysis

Details

Title: Subtitle: ATP receptor regulation of adenylate cyclase and protein kinase C activity in cultured renal LLC-PK1 cells
Creators: Robert J Anderson - Medical Service, Denver Veterans Affairs Medical Center, Colorado 80220
Ruth Breckon
Bradley S Dixon
Resource Type: Journal article
Publication Details: The Journal of clinical investigation, Vol.87(5), pp.1732-1738
DOI: 10.1172/JCI115191
PMID: 1850760
PMCID: PMC295279
ISSN: 0021-9738
eISSN: 1558-8238
Language: English
Date published: 05/1991
Academic Unit: Nephrology; Internal Medicine
Record Identifier: 9984094715702771

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