title: Structure-function relationships of molluscan troponin T revealed by limited proteolysis.
abstract: Molluscan troponin regulates muscle contraction through a novel Ca(2+)-dependent activating mechanism associated with Ca(2+)-binding to the C-terminal domain of troponin C. To elucidate the further details of this regulation, we performed limited chymotryptic digestion of the troponin complex from akazara scallop striated muscle. The results indicated that troponin T is very susceptible to the protease, compared to troponin C or troponin I. The cleavage occurred at the C-terminal extension, producing an N-terminal 33-kDa fragment and a C-terminal 6-kDa fragment. This extension is conserved in various invertebrate troponin T proteins, but not in vertebrate troponin T. A ternary complex composed of the 33-kDa fragment of troponin T, troponin I, and troponin C could be separated from the 6-kDa troponin T fragment by gel filtration. This complex did not show any Ca(2+)-dependent activation of the Mg-ATPase activity of rabbit-actomyosin-scallop-tropomyosin. In addition, the actin-tropomyosin-binding affinity of this complex was significantly decreased with increasing Ca(2+) concentration. These results indicate that the C-terminal extension of molluscan troponin T plays a role in anchoring the troponin complex to actin-tropomyosin filaments and is essential for regulation.
authors.affiliation: Laboratory of Marine Biotechnology and Microbiology, Graduate School of Fisheries Sciences, Hokkaido University, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Hisoka
authors.initials: H
authors.lastname: Suzuki
authors.forename: Iwao
authors.initials: I
authors.lastname: Ohtsuki
authors.forename: Takao
authors.initials: T
authors.lastname: Ojima
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002918:Chymotrypsin / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008969:Molecular Sequence Data; D049897:Pectinidae / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D020107:Troponin T / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*
chemical_list: D020107:Troponin T; D002918:Chymotrypsin