Binding of calcium and magnesium to human cardiac troponin C
| Autor: | R. John Solaro, Alison Yueh Li, Steffen Lindert, Filip Van Petegem, Kaveh Rayani, Anne M. Spuches, Jonathan P. Davis, Glen F. Tibbits, Justin T Seffernick |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
thermodynamic integration TF thin filament myofilament Regulatory site cTn cardiac troponin Biochemistry Troponin complex Troponin I Magnesium education.field_of_study CaM calmodulin biology Chemistry ITC isothermal titration calorimetry Cardiac muscle TI thermodynamic integration musculoskeletal system cTnC cardiac troponin C medicine.anatomical_structure Thermodynamics sTnC skeletal muscle troponin TnC calorimetry Protein Binding Research Article Calmodulin Cations Divalent Population contractility cTnI cardiac troponin I 03 medical and health sciences PDB Protein Data Bank medicine cTnT cardiac troponin T Humans education Molecular Biology Actin Binding Sites 030102 biochemistry & molecular biology FF fast-flow Myocardium Isothermal titration calorimetry MD simulation Cell Biology ITC molecular dynamics 030104 developmental biology Biophysics biology.protein Calcium Troponin C |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 1083-351X |
| Popis: | Cardiac muscle thin filaments are composed of actin, tropomyosin, and troponin that change conformation in response to Ca2+ binding, triggering muscle contraction. Human cardiac troponin C (cTnC) is the Ca2+-sensing component of the thin filament. It contains structural sites (III/IV) that bind both Ca2+ and Mg2+ and a regulatory site (II) that has been thought to bind only Ca2+. Binding of Ca2+ at this site initiates a series of conformational changes that culminate in force production. However, the mechanisms that underpin the regulation of binding at site II remain unclear. Here, we have quantified the interaction between site II and Ca2+/Mg2+ through isothermal titration calorimetry and thermodynamic integration simulations. Direct and competitive binding titrations with WT N-terminal cTnC and full-length cTnC indicate that physiologically relevant concentrations of both Ca2+/Mg2+ interacted with the same locus. Moreover, the D67A/D73A N-terminal cTnC construct in which two coordinating residues within site II were removed was found to have significantly reduced affinity for both cations. In addition, 1 mM Mg2+ caused a 1.4-fold lower affinity for Ca2+. These experiments strongly suggest that cytosolic-free Mg2+ occupies a significant population of the available site II. Interaction of Mg2+ with site II of cTnC likely has important functional consequences for the heart both at baseline as well as in diseased states that decrease or increase the availability of Mg2+, such as secondary hyperparathyroidism or ischemia, respectively. |
| Databáze: | OpenAIRE |
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