Atomic resolution probe for allostery in the regulatory thin filament

Michael R. Williams, Sarah J. Lehman, Jil C. Tardiff, Steven D. Schwartz

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

Calcium binding and dissociation within the cardiac thin filament (CTF) is a fundamental regulator of normal contraction and relaxation. Although the disruption of this complex, allosterically mediated process has long been implicated in human disease, the precise atomiclevel mechanisms remain opaque, greatly hampering the development of novel targeted therapies. To address this question, we used a fully atomistic CTF model to test both Ca2+ binding strength and the energy required to remove Ca2+ from the N-lobe binding site in WT and mutant troponin complexes that have been linked to genetic cardiomyopathies. This computational approach is combined with measurements of in vitro Ca2+ dissociation rates in fully reconstituted WT and cardiac troponin T R92L and R92W thin filaments. These human disease mutations represent known substitutions at the same residue, reside at a significant distance from the calcium binding site in cardiac troponin C, and do not affect either the binding pocket affinity or EF-hand structure of the binding domain. Both have been shown to have significantly different effects on cardiac function in vivo. We now show that these mutations independently alter the interaction between the Ca2+ ion and cardiac troponin I subunit. This interaction is a previously unidentified mechanism, in which mutations in one protein of a complex indirectly affect a third via structural and dynamic changes in a second to yield a pathogenic change in thin filament function that results in mutation-specific disease states. We can now provide atom-level insight that is potentially highly actionable in drug design.

Original languageEnglish (US)
Pages (from-to)3257-3262
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number12
DOIs
StatePublished - Mar 22 2016

Keywords

  • Calcium homeostasis
  • Cardiac thin filament
  • Hypertrophic cardiomyopathy
  • Molecular modeling
  • Steered molecular dynamics

ASJC Scopus subject areas

  • General

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