Calponin is not phosphorylated during contractions of porcine carotid arteries.
Calponin and caldesmon were purified from porcine carotid arteries that were preincubated with [32P]orthophosphate, and the stoichiometry of phosphorylation was measured. In resting arteries, caldesmon was phosphorylated to a level of 0.41 mol PO4/mol protein, while calponin was phosphorylated to levels < 0.01 mol PO4/mol protein. Stimulation by histamine (1 or 5 min), KCl (1, 5 or 60 min), or phorbol 12,13-dibutyrate (PDBu; 1 microM for 15 or 60 min) did not lead to measurable increases in the PO4 content of calponin. Because dephosphorylation of calponin during the purification procedure could account for these results, we also determined stoichiometries after firat denaturing endogenous phosphatases with trichloroacetic acid. In these experiments, calponin was determined to be phosphorylated to the same low levels as in the first set of experiments. Collectively, these data show that calponin is not phosphorylated to significant levels during contractions of carotid arteries under conditions where caldesmon phosphorylation is apparent. The circumstances under which calponin may be phosphorylated in intact smooth muscle, and the purpose that may be served by this potential regulatory process, remain to be determined.
Adam
L P
LP
Department of Medicine, Indiana University School of Medicine, Indianapolis 46202-4800, USA.
Haeberle
J R
JR
Hathaway
D R
DR
D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D002339:Carotid Arteries / Q000502:physiology*; D004988:Ethers, Cyclic / Q000494:pharmacology; D008840:Microfilament Proteins; D009124:Muscle Proteins / Q000378:metabolism; D019319:Okadaic Acid; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D010766:Phosphorylation; D013552:Swine; D014661:Vasoconstriction / Q000502:physiology*; D000096985:Calponins
D002135:Calcium-Binding Proteins; D002148:Calmodulin-Binding Proteins; D004988:Ethers, Cyclic; D008840:Microfilament Proteins; D009124:Muscle Proteins; D019319:Okadaic Acid; D015240:Phorbol 12,13-Dibutyrate; D010749:Phosphoprotein Phosphatases; D000096985:Calponins