Characterization of caldesmon binding to myosin.
Caldesmon inhibits the binding of skeletal muscle subfragment-1 (S-1).ATP to actin but enhances the binding of smooth muscle heavy meromyosin (HMM).ATP to actin. This effect results from the direct binding of caldesmon to myosin in the order of affinity: smooth muscle HMM greater than skeletal muscle HMM greater than smooth muscle S-1 greater than skeletal muscle S-1 (Hemric, M. E., and Chalovich, J. M. (1988) J. Biol. Chem. 263, 1878-1885). We now show that the difference between skeletal muscle HMM and S-1 is due to the presence of the S-2 region in HMM and is unrelated to light chain composition or to two-headed versus single-headed binding. Differences between the binding of smooth and skeletal muscle myosin subfragments to actin do not result from the lack of light chain 2 in skeletal muscle S-1. In the presence of ATP, caldesmon binds to smooth muscle myosin filaments with a stoichiometry of 1:1 (K = 1 x 10(6) M-1). Similar results were obtained for the binding of caldesmon to smooth muscle rod as well as the binding of the purified myosin-binding fragment of caldesmon to smooth muscle myosin. The binding of caldesmon to intact myosin is ATP sensitive. The interaction of caldesmon with myosin is apparently specific and sensitive to the structure of both proteins.
Hemric
M E
ME
Department of Biochemistry, East Carolina University School of Medicine, Greenville, North Carolina 27858.
Chalovich
J M
JM
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002148:Calmodulin-Binding Proteins / Q000378:metabolism*; D002645:Chickens; D002918:Chymotrypsin / Q000378:metabolism; D005895:Gizzard, Avian / Q000737:chemistry; D007460:Iodoacetamide / Q000378:metabolism; D009130:Muscle, Smooth / Q000737:chemistry; D009132:Muscles / Q000737:chemistry; D015879:Myosin Subfragments / Q000378:metabolism; D009994:Osmolar Concentration; D010206:Papain / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism
D002148:Calmodulin-Binding Proteins; D015879:Myosin Subfragments; D010446:Peptide Fragments; D000255:Adenosine Triphosphate; D002918:Chymotrypsin; D010206:Papain; D007460:Iodoacetamide