Phosphorylation by protein kinase A enhances delayed rectifier K+ current in rabbit vascular smooth muscle cells

• Published in: The American journal of physiology Vol. 268; no. 2 Pt 2; p. H926
• Main Authors: Aiello, E A, Walsh, M P, Cole, W C
• Format: Journal Article
• Language: English
• Published: United States 01.02.1995
• Subjects: 4-Aminopyridine - pharmacology; Adenylyl Imidodiphosphate - pharmacology; Animals; Cell Separation; Colforsin - pharmacology; Cyclic AMP-Dependent Protein Kinases - antagonists & inhibitors; Cyclic AMP-Dependent Protein Kinases - metabolism; Isoproterenol - pharmacology; Muscle, Smooth, Vascular - cytology; Muscle, Smooth, Vascular - physiology; Phosphorylation; Potassium - physiology; Rabbits
• ISSN: 0002-9513
• DOI: 10.1152/ajpheart.1995.268.2.h926

The effect of adenosine 3',5'-cyclic monophosphate-dependent protein kinase (PKA) activity on 4-aminopyridine (4-AP)-sensitive delayed rectifier current (IdK) in isolated rabbit portal vein smooth muscle cells was studied via whole cell voltage clamp (20-22 degrees C). A threefold increase in 4-AP-sensitive (5 mM) IdK was recorded after gaining cell access during dialysis with 5 mM intracellular ATP and internal Ca2+ buffered to a low level with 5 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. Dialysis with the nonhydrolyzable ATP analogue 5'-adenylylimidodiphosphate (5 mM) or the specific peptide inhibitor of PKA (PKI; 10 microM) reduced current runup by 50 and 70%, respectively. Delayed dialysis with PKI reversed runup and inhibited IdK to below initial levels. Forskolin (1 microM) caused a reversible increase in IdK, which was inhibited by 4-AP (5 mM). Isoproterenol (1 microM) reversibly enhanced IdK; the increase was sensitive to propranolol (2 microM) and 4-AP (5 mM) and was prevented by dialysis with PKI (10 microM). IdK was enhanced over the entire voltage range of activation and associated with a negative shift in reversal potential of net whole cell current, consistent with hyperpolarization of resting membrane potential. The data provide the first evidence for a signal transduction mechanism involving beta-adrenoceptors, adenylate cyclase, and a phosphotransferase reaction mediated by PKA for the regulation of delayed rectifier K+ channels in vascular smooth muscle.