Chemistry

A novel mutant cardiac troponin C disrupts molecular motions critical for calcium binding affinity and cardiomyocyte contractility

Document Type

Article

Abstract

Troponin C (TnC) belongs to the superfamily of EF-hand (helix-loop-helix) Ca2+-binding proteins and is an essential component of the regulatory thin filament complex. In a patient diagnosed with idiopathic dilated cardiomyopathy, we identified two novel missense mutations localized in the regulatory Ca2+-binding Site II of TnC, TnC(E59D,D75Y). Expression of recombinant TnC(E59D,D75Y) in isolated rat cardiomyocytes induced a marked decrease in contractility despite normal intracellular calcium homeostasis in intact cardiomyocytes and resulted in impaired myofilament calcium responsiveness in Triton- permeabilized cardiomyocytes. Expression of the individual mutants in cardiomyocytes showed that TnCD75Y was able to recapitulate the TnC(E59D,D75Y) phenotype, whereas TnCE59D was functionally benign. Force-pCa relationships in TnC(E59D,D75Y) reconstituted rabbit psoas fibers and fluorescence spectroscopy of TnC(E59D,D75Y) labeled with 2-[(4'-iodoacetamide)-aniline]-naphthalene-6-sulfonic acid showed a decrease in myofilament Ca2+ sensitivity and Ca2+ binding affinity, respectively. Furthermore, computational analysis of TnC showed the Ca (E59D,D75Y)-binding pocket as an active region of concerted motions, which are decreased markedly by mutation D75Y. We conclude that D75Y interferes with proper concerted motions within the regulatory Ca2+-binding pocket of TnC that hinders the relay of the thin filament calcium signal, thereby providing a primary stimulus for impaired cardiomyocyte contractility. This in turn may trigger pathways leading to aberrant ventricular remodeling and ultimately a dilated cardiomyopathy phenotype. © 2008 by the Biophysical Society.

Publication Title

Biophysical Journal

Publication Date

5-2008

Volume

94

Issue

9

First Page

3577

Last Page

3589

ISSN

0006-3495

DOI

10.1529/biophysj.107.112896

Cross Post Location

Student Publications

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