Abstract
Hypertrophic cardiomyopathy (HCM) is the most common inherited cardiovascular disease characterized by thickening of ventricular walls and decreased left ventricular chamber volume. The majority of HCM-associated mutations are found in genes encoding sarcomere proteins. Herein, we set out to functionally characterize a novel HCMassociated mutation (K206I-TNNI3) and elucidate the mechanism of dysfunction at the level of myofilament proteins. Methods and Results-The male index case was diagnosed with HCM after an out-of-hospital cardiac arrest, which was followed by comprehensive clinical evaluation, transthoracic echocardiography, and clinical genetic testing. To determine molecular mechanism(s) of the mutant human cardiac troponin I (K206I), we tested the Ca2+ dependence of thin filament-activated myosin-S1-ATPase activity in a reconstituted, regulated, actomyosin system comparing wild-type human troponin complex, 50% mix of K206I/wildtype, or 100% K206I. We also exchanged native troponin detergent extracted fibers with reconstituted troponin containing either wildtype or a 65% mix of K206I/wildtype and measured force generation. The Ca2+ sensitivity of the myofilaments containing the K206I variant was significantly increased, and when treated with 20 ìmol/L (-)-epigallocatechin gallate (green tea) was restored back to wild-type levels in ATPase and force measurements. The K206I mutation impairs the ability of the troponin I to inhibit ATPase activity in the absence of calcium-bound human cardiac troponin C. The ability of calcium-bound human cardiac troponin C to neutralize the inhibition of K206I was greater than with wild-type TnI. Conclusions-Compromised interactions of K206I with actin and hcTnC may lead to impaired relaxation and HCM.
Original language | English (US) |
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Pages (from-to) | 765-773 |
Number of pages | 9 |
Journal | Circulation: Cardiovascular Genetics |
Volume | 8 |
Issue number | 6 |
DOIs | |
State | Published - Dec 1 2015 |
Keywords
- Actin
- actomyosin
- calcium
- echocardiography
- troponin
ASJC Scopus subject areas
- Genetics
- Cardiology and Cardiovascular Medicine
- Genetics(clinical)