Single molecule kinetics in the familial hypertrophic cardiomyopathy D166V mutant mouse heart.

Abstract

One of the sarcomeric mutations associated with a malignant phenotype of familial hypertrophic cardiomyopathy (FHC) is the D166V point mutation in the ventricular myosin regulatory light chain (RLC) encoded by the MYL2 gene. In this report we show that the rates of myosin cross-bridge attachment and dissociation are significantly different in isometrically contracting cardiac myofibrils from right ventricles of transgenic (Tg)-D166V and Tg-WT mice. We have derived the myosin cross-bridge kinetic rates by tracking the orientation of a fluorescently labeled single actin molecule. Orientation (measured by polarized fluorescence) oscillated between two states, corresponding to the actin-bound and actin-free states of the myosin cross-bridge. The rate of cross-bridge attachment during isometric contraction decreased from 3 s(-1) in myofibrils from Tg-WT to 1.4 s(-1) in myofibrils from Tg-D166V. The rate of detachment decreased from 1.3 s(-1) (Tg-WT) to 1.2 s(-1) (Tg-D166V). We also showed that the level of RLC phosphorylation was largely decreased in Tg-D166V myofibrils compared to Tg-WT. Our findings suggest that alterations in the myosin cross-bridge kinetics brought about by the D166V mutation in RLC might be responsible for the compromised function of the mutated hearts and lead to their inability to efficiently pump blood.

• Authors: Priya Muthu, Prasad Mettikolla, Nils Calander, Rafal Luchowski, Ignacy Gryczynski, Zygmunt Gryczynski, Danuta Szczesna-Cordary, J Borejdo
• Journal: Journal of molecular and cellular cardiology (J Mol Cell Cardiol) Vol. 48 Issue 5 Pg. 989-98 (May 2010) ISSN: 1095-8584 [Electronic] England
• PMID: 19914255 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright (c) 2009 Elsevier Ltd. All rights reserved.

Chemical References

• Myosin Light Chains

Topics

• Animals
• Anisotropy
• Cardiomyopathy, Hypertrophic, Familial (metabolism)
• In Vitro Techniques
• Kinetics
• Mice
• Mice, Transgenic
• Myofibrils (metabolism)
• Myosin Light Chains (metabolism)
• Phosphorylation
• Sarcomeres (metabolism)
• Spectrometry, Fluorescence