Abstract
On treatment with 10mM EDTA at 30°C, protein of 18, 000 daltons was released from myofibrils, thin filaments and myosin B prepared from the smooth muscle of an ascidian, Halocynthia roretzi. This protein was purified from the EDTA extract of myofibrils by differential centrifugation, freeze-drying and gel-filtration. Based on its molecular weight, electrophoretic mobilities in the presence and absence of Ca2+ and other properties, it was identified as troponin C.
By EDTA treatment, ascidian myosin B lost the Ca2+-sensitivity of Mg2+-ATPase, and EDTA-treated myosin B recovered the sensitivity by mixing with the EDTA extract of myosin B in the presence of Mg2+. Gel-electrophoretic patterns indicated that desensitization and resensitization of ascidian myosin B were accom-panied by the removal and binding of troponin C. These results indicate that ascidian smooth muscle is regulated by a troponin-tropomyosin system, and desensitization induced by EDTA treatment is due to the removal of troponin C but not the release of the light chains of the myosin molecule.
Based on these findings, we have established a simple method for the purification of troponin C from ascidian smooth muscle.
