title: Calcium-induced peptide association to form an intact protein domain: 1H NMR structural evidence.
abstract: The 70-residue carboxyl-terminal domain of the muscle contractile protein troponin-C contains two helix-loop-helix calcium (Ca)-binding sites that are related to each other by approximate twofold rotational symmetry. Hydrophobic residues from the helices and a short three residue beta sheet at the interface of the two sites act to stabilize the protein domain in the presence of Ca. A synthetic 34-residue peptide representing one of these sites (site III) has been synthesized and studied by H-1 nuclear magnetic resonance (NMR) spectroscopy. In solution this peptide undergoes a Ca-induced conformational change to form the helix-loop-helix Ca-binding motif. Two-dimensional nuclear Overhauser effect spectra have provided evidence for the formation of a beta sheet and interactions between several hydrophobic residues from opposing helices as found in troponin-C. It is proposed that a symmetric two-site dimer similar in tertiary structure to the carboxyl-terminal domain of troponin-C forms from the assembly of two site III peptides in the Ca-bound form.
authors.affiliation: Department of Biochemistry, University of Alberta, Edmonton, Canada.
authors.forename: G S
authors.initials: GS
authors.lastname: Shaw
authors.forename: R S
authors.initials: RS
authors.lastname: Hodges
authors.forename: B D
authors.initials: BD
authors.lastname: Sykes
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D006859:Hydrogen; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism*; D011487:Protein Conformation; D014336:Troponin / Q000138:chemical synthesis / Q000378:metabolism*; D019209:Troponin C; D014422:Turkeys
chemical_list: D010455:Peptides; D014336:Troponin; D019209:Troponin C; D006859:Hydrogen; D002118:Calcium