Effects of removal and reconstitution of myosin regulatory light chain and troponin C on the Ca(2+)-sensitive ATPase activity of myofibrils from scallop striated muscle.
In order to examine the involvement of troponin-linked Ca(2+)-regulation, in addition to well-known myosin-linked Ca(2+)-regulation, in the contraction of molluscan striated muscle, myofibrils from Ezo-giant scallop striated muscle were desensitized to Ca(2+) by removing both myosin regulatory light chain and troponin C by treatment with a strong divalent cation chelator, CDTA. The ATPase level in the desensitized myofibrils was about half the maximum level in intact myofibrils regardless of the Ca(2+)-concentration at 25 and 15 degrees C. In the absence of Ca(2+), the ATPase of the desensitized myofibrils was suppressed by myosin regulatory light chain but not affected by troponin C at either temperature. The ATPase was activated at higher Ca(2+)-concentrations by both myosin regulatory light chain and troponin C, but the activating effects of these two proteins were affected differently by temperature. The activation of ATPase by myosin regulatory light chain was much greater than that by troponin C at 25 degrees C, whereas the activation by troponin C was much greater than that by myosin regulatory light chain at 15 degrees C. The maximum activation was only obtained in the presence of both myosin regulatory light chain and troponin C at these temperatures. These findings strongly suggest that the contraction of scallop striated muscle is regulated through both myosin-linked and troponin-linked Ca(2+)-regulation, and that the troponin-linked Ca(2+)-regulation is more significant at lower temperature.
Shiraishi
F
F
Department of Pharmacology, Faculty of Medicine, Kyushu University, Fukuoka, 812-8582, Japan.
Morimoto
S
S
Nishita
K
K
Ojima
T
T
Ohtsuki
I
I
D000818:Animals; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D003080:Cold Temperature; D004591:Electrophoresis, Polyacrylamide Gel; D008974:Mollusca; D018482:Muscle, Skeletal / Q000201:enzymology*; D009210:Myofibrils / Q000201:enzymology*; D018994:Myosin Light Chains / Q000378:metabolism*; D019209:Troponin C / Q000378:metabolism*
D018994:Myosin Light Chains; D019209:Troponin C; D000252:Calcium-Transporting ATPases