title: NH2-terminal truncation of skeletal muscle troponin T does not alter the Ca2+ sensitivity of thin filament assembly.
abstract: To investigate how Ca2+ binding to troponin C regulates muscle contraction, the Ca(2+)-sensitive properties of thin filament assembly were studied as the tropomyosin binding, NH2-terminal region of troponin T was progressively shortened. Troponin complexes were prepared that contained skeletal muscle troponin C, troponin I, and either intact troponin T (TnT) (residues 1-259) or fragment TnT-(70-259), TnT-(151-259), or TnT-(159-259). In the absence of Ca2+ their respective affinities for pyrene-labeled tropomyosin were 2.3 x 10(7) M-1, 1.2 x 10(7) M-1, 1.9 x 10(5) M-1, and 1.9 x 10(5) M-1. Ca2+ had only a small effect on these affinities: 1.1 x 10(7) M-1 for whole troponin, 2 x 10(5) M-1 for troponin-(151-259), and 2.8 x 10(5) M-1 for troponin-(159-259). Forms of troponin that bound weakly to tropomyosin in the absence of actin increased the actin affinity of tropomyosin only 2-3-fold, even in the absence of Ca2+; weak binding of troponin to tropomyosin correlated with weak effects on tropomyosin-actin binding. In contrast, whole troponin had an approximately 500-fold effect on tropomyosin binding to actin, regardless of whether Ca2+ was present. The small effect of Ca2+ on the energetics of thin filament assembly is not attributable to the amino-terminal region of troponin T. The results suggest that Ca2+ causes the interaction between actin and the globular region of troponin to switch between two energetically similar states.
authors.affiliation: Department of Biochemistry, University of Iowa, Iowa City 52242, USA.
authors.forename: D
authors.initials: D
authors.lastname: Fisher
authors.forename: G
authors.initials: G
authors.lastname: Wang
authors.forename: L S
authors.initials: LS
authors.lastname: Tobacman
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D000199:Actins / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D004252:DNA Mutational Analysis; D008954:Models, Biological; D018482:Muscle, Skeletal / Q000737:chemistry*; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits; D011994:Recombinant Proteins / Q000378:metabolism; D017384:Sequence Deletion; D014335:Tropomyosin / Q000378:metabolism; D014336:Troponin / Q000235:genetics / Q000378:metabolism*; D020107:Troponin T
chemical_list: D000199:Actins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014335:Tropomyosin; D014336:Troponin; D020107:Troponin T; D002118:Calcium