Conformation of the regulatory domain of cardiac muscle troponin C in its complex with cardiac troponin I.
Calcium activation of fast striated muscle results from an opening of the regulatory N-terminal domain of fast skeletal troponin C (fsTnC), and a substantial exposure of a hydrophobic patch, essential for Ca(2+)-dependent interaction with fast skeletal troponin I (fsTnI). This interaction is obligatory to relieve the inhibition of strong, force-generating actin-myosin interactions. We have determined intersite distances in the N-terminal domain of cardiac TnC (cTnC) by fluorescence resonance energy transfer measurements and found negligible increases in these distances when the single regulatory site is saturated with Ca(2+). However, in the presence of bound cardiac TnI (cTnI), activator Ca(2+) induces significant increases in the distances and a substantial opening of the N-domain. This open conformation within the cTnC.cTnI complex has properties favorable for the Ca(2+)-induced interaction with an additional segment of cTnI. Thus, the binding of cTnI to cTnC is a prerequisite to achieve a Ca(2+)-induced open N-domain similar to that previously observed in fsTnC with no bound fsTnI. This role of cardiac TnI has not been previously recognized. Our results also indicate that structural information derived from a single protein may not be sufficient for inference of a structure/function relationship.
Dong
W J
WJ
Department of Biochemistry, University of Alabama at Birmingham, Birmingham, Alabama 35294-2041, USA.
Xing
J
J
Villain
M
M
Hellinger
M
M
Robinson
J M
JM
Chandra
M
M
Solaro
R J
RJ
Umeda
P K
PK
Cheung
H C
HC
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002642:Chick Embryo; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D008958:Models, Molecular; D009154:Mutation; D009206:Myocardium / Q000737:chemistry*; D009282:Naphthalenesulfonates; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D019209:Troponin C / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019210:Troponin I / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014364:Tryptophan / Q000737:chemistry
D005456:Fluorescent Dyes; D009282:Naphthalenesulfonates; D010446:Peptide Fragments; D011994:Recombinant Proteins; D019209:Troponin C; D019210:Troponin I; C006421:1,5-I-AEDANS; D014364:Tryptophan; D002118:Calcium