Changes in the dynamics of the cardiac troponin C molecule explain the effects of Ca2+-sensitizing mutations
| Autor: | Gurpreet Singh, Charles M. Stevens, D. Peter Tieleman, Bairam Lotfalisalmasi, Kaveh Rayani, Glen F. Tibbits |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Regulation of gene expression chemistry.chemical_classification Myofilament 030102 biochemistry & molecular biology biology Chemistry Mutant Wild type Isothermal titration calorimetry Peptide Cell Biology musculoskeletal system Biochemistry Troponin 03 medical and health sciences 030104 developmental biology Troponin I cardiovascular system biology.protein Biophysics Molecular Biology |
| Zdroj: | Journal of Biological Chemistry. 292:11915-11926 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m116.770776 |
| Popis: | Cardiac troponin C (cTnC) is the regulatory protein that initiates cardiac contraction in response to Ca2+. TnC binding Ca2+ initiates a cascade of protein–protein interactions that begins with the opening of the N-terminal domain of cTnC, followed by cTnC binding the troponin I switch peptide (TnISW). We have evaluated, through isothermal titration calorimetry and molecular-dynamics simulation, the effect of several clinically relevant mutations (A8V, L29Q, A31S, L48Q, Q50R, and C84Y) on the Ca2+ affinity, structural dynamics, and calculated interaction strengths between cTnC and each of Ca2+ and TnISW. Surprisingly the Ca2+ affinity measured by isothermal titration calorimetry was only significantly affected by half of these mutations including L48Q, which had a 10-fold higher affinity than WT, and the Q50R and C84Y mutants, each of which had affinities 3-fold higher than wild type. This suggests that Ca2+ affinity of the N-terminal domain of cTnC in isolation is insufficient to explain the pathogenicity of these mutations. Molecular-dynamics simulation was used to evaluate the effects of these mutations on Ca2+ binding, structural dynamics, and TnI interaction independently. Many of the mutations had a pronounced effect on the balance between the open and closed conformations of the TnC molecule, which provides an indirect mechanism for their pathogenic properties. Our data demonstrate that the structural dynamics of the cTnC molecule are key in determining myofilament Ca2+ sensitivity. Our data further suggest that modulation of the structural dynamics is the underlying molecular mechanism for many disease mutations that are far from the regulatory Ca2+-binding site of cTnC. |
| Databáze: | OpenAIRE |
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