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J Biol Chem, Vol. 273, Issue 8, 4695-4704, February 20, 1998

Coexpression of Ligand-gated P2X and G Protein-coupled P2Y Receptors in Smooth Muscle
PREFERENTIAL ACTIVATION OF P2Y RECEPTORS COUPLED TO PHOSPHOLIPASE C (PLC)-beta 1 VIA Galpha q/11 AND TO PLC-beta 3 VIA Gbeta gamma i3

Karnam S. Murthy and Gabriel M. Makhlouf

From the Departments of Medicine and Physiology, Medical College of Virginia, Richmond, Virginia 23298-0711

P2 receptor subtypes and their signaling mechanisms were characterized in dispersed smooth muscle cells. UTP and ATP stimulated inositol 1,4,5-triphosphate formation, Ca2+ release, and contraction that were abolished by U-73122 and guanosine 5'-O-(3-thio)diphosphate, and partly inhibited (50-60%) by pertussis toxin (PTX). ATP analogs (adenosine 5'-(alpha ,beta -methylene)triphosphate, adenosine 5'-(beta ,gamma -methylene)triphosphate, and 2-methylthio-ATP) stimulated Ca2+ influx and contraction that were abolished by nifedipine and in Ca2+-free medium. Micromolar concentrations of ATP stimulated both Ca2+ influx and Ca2+ release.

ATP and UTP activated Gq/11 and Gi3 in gastric and aortic smooth muscle and heart membranes, Gq/11 and Gi1 and/or Gi2 in liver membranes, and Go and Gi1-3 in brain membranes. Phosphoinositide hydrolysis stimulated by ATP and UTP was mediated concurrently by Galpha q/11-dependent activation of phospholipase (PL) C-beta 1 and Gbeta gamma i3-dependent activation of PLC-beta 3. Phosphoinositide hydrolysis was partially inhibited by PTX or by antibodies to Galpha q/11, Gbeta , PLC-beta 1, or PLC-beta 3, and completely inhibited by the following combinations (PLC-beta 1 and PLC-beta 3 antibodies; Galpha q/11 and Gbeta antibodies; PLC-beta 1 and Gbeta antibodies; PTX with either PLC-beta 1 or Galpha q/11 antibody).

The pattern of responses implied that P2Y2 receptors in visceral, and probably vascular, smooth muscle are coupled to PLC-beta 1 via Galpha q/11 and to PLC-beta 3 via Gbeta gamma i3. These receptors co-exist with ligand-gated P2X1 receptors activated by ATP analogs and high levels of ATP.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.


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