The role of electrostatics in the interaction of the inhibitory region of troponin I with troponin C.
We have addressed the electrostatic interactions occurring between the inhibitory region of cardiac troponin I with the C-lobe of troponin C using scanning glycine mutagenesis of the inhibitory region. We report variations in the electric potentials due to mutation of charged residues within this complex based upon the solved NMR structure (1OZS). These results demonstrate the importance of electrostatics within this complex, and it is proposed that electrostatic interactions are integral to the formation and function of larger ternary troponin complexes. To address this hypothesis, we report (15)N NMR relaxation measurements, which suggest that, within a ternary complex involving the C-lobe and the N-terminal region of troponin I (residues 34-71), the inhibitory region maintains the electrostatic interactions with the E-helix of the C-lobe as observed within the binary complex. These results imply that, in solution, the cardiac troponin complex behaves in a manner consistent with that of the crystal structure of the skeletal isoform (1YTZ). A cardiac troponin complex possessing domain orientations similar to that of the skeletal isoform provides structural insights into altered troponin I activities as observed for the familial hypertrophic cardiomyopathy mutation R144G and phosphorylation of Thr142.
Lindhout
Darrin A
DA
CIHR Group in Protein Structure and Function, Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2H7.
Boyko
Robert F
RF
Corson
David C
DC
Li
Monica X
MX
Sykes
Brian D
BD
D000595:Amino Acid Sequence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry; D055672:Static Electricity; D013816:Thermodynamics; D019209:Troponin C / Q000737:chemistry* / Q000378:metabolism; D019210:Troponin I / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D010455:Peptides; D019209:Troponin C; D019210:Troponin I