Tyrosine phosphorylation and regulation of swine carotid artery contraction.
Regulation of [Ca2+]i and modulation of the [Ca2+]i sensitivity of myosin phosphorylation in smooth muscles may involve phosphorylation of one or more proteins on tyrosine residues. We tested this hypothesis by measuring tyrosine phosphorylation of proteins extracted from swine carotid artery and separated by SDS gel electrophoresis. Tyrosine phosphorylation was estimated by the binding of antiphosphotyrosine antibodies to proteins in SDS gels. We found four bands with approximate molecular weights of 120, 110, 85 and 75 kD in which tyrosine phosphorylation increased 1 min after histamine stimulation. After washout of histamine, dephosphorylation of the proteins in these four bands occurred at a slower rate than relaxation. Tyrosine phosphorylation of these four protein bands did not correlate with the [Ca2+]i sensitivity of myosin phosphorylation following agonist or high [K+]o stimulation. Phorbol dibutyrate stimulation also induced tyrosine phosphorylation of these four protein bands. These data suggest that tyrosine phosphorylation of the proteins in these four bands may be involved in the initial phase of swine carotid artery contraction. However, there was, at most, only a minor involvement of tyrosine phosphorylation of these four protein bands in the sustained phase of contraction or in the regulation of [Ca2+]i sensitivity of myosin phosphorylation in the swine carotid artery. These data do not rule out a role for other, less abundant tyrosine phosphoproteins in the regulation of sustained contraction or the [Ca2+]i sensitivity of myosin phosphorylation.
Rembold
C M
CM
Department of Internal Medicine, University of Virginia Health Sciences Center, Charlottesville 22908, USA.
Weaver
B A
BA
D000818:Animals; D002118:Calcium / Q000502:physiology*; D002339:Carotid Arteries / Q000502:physiology*; D009119:Muscle Contraction / Q000502:physiology*; D010766:Phosphorylation; D013552:Swine; D014443:Tyrosine / Q000378:metabolism*
D014443:Tyrosine; D002118:Calcium