title: Insights into human beta-cardiac myosin function from single molecule and single cell studies.
abstract: beta-Cardiac myosin is a mechanoenzyme that converts the energy from ATP hydrolysis into a mechanical force that drives contractility in muscle. Thirty percent of the point mutations that result in hypertrophic cardiomyopathy are localized to MYH7, the gene encoding human beta-cardiac myosin heavy chain (beta-MyHC). Force generation by myosins requires a tight and highly conserved allosteric coupling between its different protein domains. Hence, the effects of single point mutations on the force generation and kinetics of beta-cardiac myosin molecules cannot be predicted directly from their location within the protein structure. Great insight would be gained from understanding the link between the functional defect in the myosin protein and the clinical phenotypes of patients expressing them. Over the last decade, several single molecule techniques have been developed to understand in detail the chemomechanical cycle of different myosins. In this review, we highlight the single molecule techniques that can be used to assess the effect of point mutations on beta-cardiac myosin function. Recent bioengineering advances have enabled the micromanipulation of single cardiomyocyte cells to characterize their force-length dynamics. Here, we briefly review single cell micromanipulation as an approach to determine the effect of beta-MyHC mutations on cardiomyocyte function. Finally, we examine the technical challenges specific to studying beta-cardiac myosin function both using single molecule and single cell approaches.
authors.affiliation: Department of Biochemistry, Stanford University, Stanford, CA, USA.
authors.forename: Sivaraj
authors.initials: S
authors.lastname: Sivaramakrishnan
authors.forename: Euan
authors.initials: E
authors.lastname: Ashley
authors.forename: Leslie
authors.initials: L
authors.lastname: Leinwand
authors.forename: James A
authors.initials: JA
authors.lastname: Spudich
mesh_terms: D000818:Animals; D001681:Biological Assay; D002312:Cardiomyopathy, Hypertrophic / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D024741:Cardiomyopathy, Hypertrophic, Familial / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D020022:Genetic Predisposition to Disease; D006801:Humans; D057054:Molecular Imaging; D009154:Mutation; D009200:Myocardial Contraction / Q000235:genetics; D032383:Myocytes, Cardiac / Q000201:enzymology* / Q000473:pathology; D010641:Phenotype; D024723:Ventricular Myosins / Q000235:genetics / Q000378:metabolism*
chemical_list: D024723:Ventricular Myosins