title: Interaction of tropomyosin and troponin T: a proton nuclear magnetic resonance study.
abstract: Proton nuclear magnetic resonance (1H NMR) has been used to study the nature of the interaction between tropomyosin (TM) and troponin T (Tn-T). Resonances corresponding to the histidine residues in fragments of both TM and Tn-T can be resolved and assigned in the 1H NMR spectrum. Changes in the pH titration profiles of these resonances when the various fragments are mixed provide probes of the interaction sites between the proteins. Fragment T1 (residues 1-158) of Tn-T appears to interact weakly but specifically with fragments of TM in which the NH2-terminus (residues 1-10) is intact. While fragment CB2 (residues 71-151) of Tn-T interacts weakly (dissociation constant of 0.1-0.2 mM) with NH2-terminal fragments of TM, this appears to be nonspecific since the absence of residues 1-10 and 128-189 of TM does not affect the observed perturbations of the titration profiles of His-79 of CB2. Although a strong interaction between T1 and the COOH-terminal Cy2 fragment (residues 190-284) of TM has been previously demonstrated, no perturbation of His-276 of Cy2 or of His-7, -23, -29, or -36 of T1 was observed in a mixture of T1/Cy2. The pKa of His-276 was also not affected in a mixture of Cy1/Cy2 (where Cy1 is residues 1-189 of TM) but was significantly decreased in the ternary complex T1/Cy1/Cy2. The importance of residues 1-70 of Tn-T in its binding to TM is illustrated by the specificity it confers on the T1/Cy1 interaction and by the absence of His-276 perturbation in the mixture CB2/Cy1/Cy2.
mesh_terms: D006639:Histidine; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D013816:Thermodynamics; D014335:Tropomyosin / Q000378:metabolism*; D014336:Troponin / Q000378:metabolism*; D020107:Troponin T
chemical_list: D010446:Peptide Fragments; D014335:Tropomyosin; D014336:Troponin; D020107:Troponin T; D006639:Histidine