A model for the function of the bisphosphorylated heart-specific troponin-I N-terminus.
Bisphosphorylation of two adjacently located serine residues in the heart-specific N-terminus of the cTnl subunit reduces calcium affinity of the cTnC subunit. An interaction of the phosphorylation region of cTnI with acidic residues of another cTn subunit has been proposed formerly based on 31P nuclear magnetic resonance (NMR) data. A possible candidate is cTnC. Thus, an interaction model of cTnC with the bisphosphorylated cTnI N-terminus has been built using a homology model of hcTnC based on the crystal structure of tusTnC and the structure of the phosphorylation region of cTnI determined by 2D NMR. By computational search, five cluster of acidic residues of cTnC might interact with the cTnI phosphorylation region. Three sites could be excluded by 31P-NMR experiments. The two remaining sites are located in the N-terminal helix of cTnC and between calcium binding sites III and IV. Reorientation of the arginine and phosphoserine sidechains within the phosphorylation region as proposed by refined docking could explain the formerly measured changes in pKaapp values. Thus, local pKa changes might lead to the reduction of calcium affinity observed upon cTnI bisphosphorylation.
Jaquet
K
K
Medizinische Fakultät, Ruhr-Universität Bochum, Germany. Kornelia.E.Jaquet@ruhr-uni-bochum.de
Lohmann
K
K
Czisch
M
M
Holak
T
T
Gulati
J
J
Jaquet
R
R
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D003198:Computer Simulation; D008958:Models, Molecular; D009206:Myocardium / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism*; D010759:Phosphorus Isotopes; D010766:Phosphorylation; D011485:Protein Binding; D011817:Rabbits; D019209:Troponin C / Q000378:metabolism; D019210:Troponin I / Q000737:chemistry* / Q000378:metabolism*
D010446:Peptide Fragments; D010759:Phosphorus Isotopes; D019209:Troponin C; D019210:Troponin I; D002118:Calcium