American lobster troponin.
Troponin was isolated from the abdominal muscle of the American lobster (Homarus americanus) by essentially the same method as used for akazara scallop troponin [J. Biol. Chem. 261, 16749-16754 (1986)]. The thus isolated troponin together with lobster tropomyosin confers high Ca2(+)-sensitivity to rabbit reconstituted actomyosin. The troponin consists of components having Mr of about 42,000, 32,000, 30,000, and 17,000, but not the Mr 52,000-59,000 component previously reported to be present in several crustacean troponins. These troponin components were separated from each other by DEAE-Toyopearl column chromatography in the presence of 6 M urea. The Mr 17,000 component was further separated into one major and two minor components by the same chromatography, but each of them was confirmed to be a Ca2+ binding component, TnC. The Mr 32,000 and 30,000 components were both regarded as inhibitory subunits, TnIs, since the Mg-ATPase activity of actomyosin in the presence of tropomyosin was strongly inhibited by the addition of the components, and the inhibition was reversed by the further addition of TnC. Finally, the Mr 42,000 component was regarded as TnT, since this component formed stoichiometic complex with TnC and TnI, and was indispensable for Ca2+ regulation of the actomyosin-tropomyosin system.
Nishita
K
K
Department of Chemistry, Faculty of Fisheries, Hokkaido University.
Ojima
T
T
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002850:Chromatography, Gel; D008970:Molecular Weight; D008121:Nephropidae / Q000032:analysis*; D012680:Sensitivity and Specificity; D014336:Troponin / Q000302:isolation & purification*
D000596:Amino Acids; D014336:Troponin; D017301:Ca(2+) Mg(2+)-ATPase; D002118:Calcium