title: Calcium-independent activation of skeletal muscle fibers by a modified form of cardiac troponin C.
abstract: A conformational change accompanying Ca2+ binding to troponin C (TnC) constitutes the initial event in contractile regulation of vertebrate striated muscle. We replaced endogenous TnC in single skinned fibers from rabbit psoas muscle with a modified form of cardiac TnC (cTnC) which, unlike native cTnC, probably contains an intramolecular disulfide bond. We found that such activating TnC (aTnC) enables force generation and shortening in the absence of calcium. With aTnC, both force and shortening velocity were the same at pCa 9.2 and pCa 4.0. aTnc could not be extracted under conditions which resulted in extraction of endogenous TnC. Thus, aTnC provides a stable model for structural studies of a calcium binding protein in the active conformation as well as a useful tool for physiological studies on the primary and secondary effects of Ca2+ on the molecular kinetics of muscle contraction.
authors.affiliation: Department of Physiology and Biophysics, University of Washington, Seattle 98195.
authors.forename: J D
authors.initials: JD
authors.lastname: Hannon
authors.forename: P B
authors.initials: PB
authors.lastname: Chase
authors.forename: D A
authors.initials: DA
authors.lastname: Martyn
authors.forename: L L
authors.initials: LL
authors.lastname: Huntsman
authors.forename: M J
authors.initials: MJ
authors.lastname: Kushmerick
authors.forename: A M
authors.initials: AM
authors.lastname: Gordon
mesh_terms: D000818:Animals; D001665:Binding Sites; D055592:Biophysical Phenomena; D001703:Biophysics; D002118:Calcium / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry; D066298:In Vitro Techniques; D009119:Muscle Contraction / Q000502:physiology*; D009132:Muscles / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry; D011817:Rabbits; D014336:Troponin / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D019209:Troponin C
chemical_list: D014336:Troponin; D019209:Troponin C; D003545:Cysteine; D002118:Calcium