CA2-DEPENDENT BINDING OF CALCYCLIN TO MUSCLE TROPOMYOSIN()

The interaction of calcyclin with tropomyosin and tropomyosin actin wa s studied with fluorescence titrations and photo-reactive crosslinking experiments. Titrations of pyreneiodoacetamide-labeled tropomyosin al one or with actin showed binding of calcyclin to tropomyosin with mu M dissociation constants when Ca2+ was present. UV irradiation of mixtu res of calcyclin and gizzard beta beta-tropomyocin labeled with benzop henone-iodoacetamide at Cys36, with or without actin, produced crossli nks between tropomyosin chains and calcyclin monomers only in the pres ence of Ca2+. These data provide direct evidence for a Ca2+-dependent tropomyosin-calcyclin interaction at or near Cys36 of tropomyosin and indicate that calcyclin binding to tropomyosin-actin does not cause tr opomyosin dissociation. (C) 1996 Academic Press, Inc.