title: Clusters of bound Ca(2+) initiate contraction in fast skeletal muscle.
abstract: Ca(2+)-binding to troponin C ultimately controls force in muscle leading to the expectation that the two curves, pCa/force and pCa/Ca(2+) binding, will coincide. Using an improved fluorescence apparatus to measure Ca(2+)-binding, we confirm a displacement between the position and shape of the pCa/Ca(2+)-binding and pCa/force curves. This displacement may be part of a mechanism that reduces the noise inherent in the control process. There must always be some Ca(2+)-binding events even at 10 or 100nM, well below threshold for muscle contraction. To minimize the response to such random binding events we suggest that clusters of adjacent Ca(2+)-binding sites must be filled before contraction is initiated. Clusters promote the reconfiguration of the thin filament to the "On" state; this simultaneously increases thin filaments' affinity for myosin heads and of troponin C for Ca(2+) producing the highly cooperative pCa/force curve. The cluster requirement displaces the Ca(2+)-binding from the force curve as observed. The thin filament conformational changes and the accompanying affinity increases introduce a discontinuity in the pCa/Ca(2+)-binding curve. The curve, therefore, is most appropriately fit by two separate Hill equations, a simple non-cooperative one (midpoint, pK1, n1∼1) for the foot and a second cooperative one (pK2, n2∼2.5) for the upper part. With this fit pK2 is larger than pK1 as our argument requires, in contrast to fitting to the sum of two Hill equations. It also expresses the idea that there may be three states of the thin filament.
authors.affiliation: Department of Pathology, Columbia University, NY, NY 10032, USA.
authors.forename: Philip W
authors.initials: PW
authors.lastname: Brandt
authors.affiliation: Dipartimento di Medicina Sperimentale e Clinica, Università degli Studi di Firenze, Firenze 50134, Italy. Electronic address: corrado.poggesi@unifi.it.
authors.forename: Corrado
authors.initials: C
authors.lastname: Poggesi
mesh_terms: D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D005453:Fluorescence; D009119:Muscle Contraction; D018656:Muscle Fibers, Fast-Twitch / Q000737:chemistry / Q000502:physiology*; D009210:Myofibrils / Q000378:metabolism; D011817:Rabbits; D019209:Troponin C / Q000378:metabolism
chemical_list: D019209:Troponin C; D002118:Calcium
keywords: Calcium binding; Contraction regulation; Fluorescent probes; Myofilaments; Skeletal muscle; Troponin C