title: A disulfide crosslink between Cys98 of troponin-C and Cys133 of troponin-I abolishes the activity of rabbit skeletal troponin.
abstract: Various thio-reactive bifunctional crosslinkers as well as 5,5'-dithiobis(2-nitrobenzoate)-mediated disulfide bond formation were used to crosslink troponin-C and troponin-I, the Ca(2+)-binding and inhibitory subunits of troponin, respectively. In all cases, substantial crosslinking was obtained when the reactions were carried out in the absence of Ca2+. No disulfide crosslinking occurred if either Cys98 of TnC, or Cys133 of TnI were blocked, indicating that these thiols are involved in the crosslinking. Troponin containing the disulfide crosslink is no longer capable of regulating actomyosin ATPase activity in a Ca(2+)-dependent manner. Our results suggest that the relative movement between the Cys98 region of TnC and the Cys133 region of TnI is required for the Ca(2+)-regulatory process in skeletal muscle.
authors.affiliation: Muscle Research Group, Boston Biomedical Research Institute, Massachusetts 02114.
authors.forename: H S
authors.initials: HS
authors.lastname: Park
authors.forename: B J
authors.initials: BJ
authors.lastname: Gong
authors.forename: T
authors.initials: T
authors.lastname: Tao
mesh_terms: D000205:Actomyosin / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D003545:Cysteine; D004220:Disulfides; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D009132:Muscles / Q000502:physiology*; D011817:Rabbits; D013997:Time Factors; D014336:Troponin / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D019210:Troponin I
chemical_list: D003432:Cross-Linking Reagents; D004220:Disulfides; D014336:Troponin; D019210:Troponin I; D000205:Actomyosin; D004228:Dithionitrobenzoic Acid; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases; D003545:Cysteine