Proton magnetic resonance studies on peptide fragments of troponin-C containing single calcium-binding sites.
Proton magnetic resonance spectroscopy has been employed to study the solution conformation of three cleavage fragments of troponin-C, each containing a single Ca(II)-binding site and corresponding to different regions in the primary sequence; viz. CB8 (residues 46-77), CB9 (residues 85-134) and TH2 (residues 121-159). Although all three peptides lack a well-defined tertiary fold in the absence of metal ions, several spectral features indicate the presence of local conformational constraints in each apo-peptide. Ca(II) binding led to spectral changes consistent with increased restriction of backbone motility and the adoption of a more compact conformation. Studies using paramagnetic ions as conformational probes support current views concerning the nature of the ligands at the metal binding sites. The nature and kinetics of the structural influence of metal binding suggest that the conformational constraints existing in the CB8 apo-peptide provide an adequate Ca(II)-binding configuration. In contrast, the CB9 and TH2 peptides exhibit spectral changes consistent with an increased local structure in the region of helix E (residues 94-102) in the case of CB9 and helix H (residues 148-159) in the case of TH2. In CB9, conformation changes also appear to be transmitted to a portion of the sequence (residues 87-93) preceding helix E, a putative site of interaction between troponin-C and troponin-I. These data are discussed with reference to the contribution of long-range (interdomain) interactions within troponin-C and the modulation of troponin subunit protein-protein interactions by Ca(II) binding.
D000818:Animals; D001665:Binding Sites; D001842:Bone and Bones / Q000032:analysis; D002118:Calcium / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D009124:Muscle Proteins / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism; D011817:Rabbits; D014336:Troponin / Q000032:analysis* / Q000378:metabolism; D019209:Troponin C
D009124:Muscle Proteins; D010446:Peptide Fragments; D014336:Troponin; D019209:Troponin C; D002118:Calcium