Single molecule detection approach to muscle study: Kinetics of a single cross-bridge during contraction of muscle

Julian Borejdo, Danuta Szczesna-Cordary, Priya Muthu, Prasad Metticolla, Rafal Luchowski, Zygmunt Gryczynski, Ignacy Gryczynski

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

6 Scopus citations

Abstract

D166V point mutation in the ventricular myosin regulatory light chain (RLC) is one of the causes of familial hypertrophic cardiomyopathy (FHC). We show here that the rates of cross-bridge attachment and dissociation are significantly different in isometrically contracting cardiac myofibrils from right ventricle of WT and Tg-D166V mice. To avoid averaging over ensembles of molecules composing muscle fibers, the data was collected from a single molecule. Kinetics were derived by tracking the orientation of a single actin molecule by fluorescence anisotropy. Orientation oscillated between two states, corresponding to the actin-bound and actin-free states of the myosin cross-bridge. The cross-bridge in a wild-type (healthy) heart stayed attached and detached from thin filament on average for 0.7 and 2.7 s, respectively. In FHC heart, these numbers increased to 2.5 and 5.8 s, respectively. These findings suggest that alterations in myosin cross-bridge kinetics associated with D166V mutation of RLC ultimately affect the ability of a heart to efficiently pump the blood.

Original languageEnglish
Title of host publicationSpectroscopic Methods of Analysis
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages311-334
Number of pages24
ISBN (Print)9781617798054
DOIs
StatePublished - 1 Jan 2012

Publication series

NameMethods in Molecular Biology
Volume875
ISSN (Print)1064-3745

Keywords

  • Cardiac muscle
  • Correlation function
  • Hearth hypertrophy
  • Single molecule detection

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