title: Functional characterization of Dictyostelium discoideum mutant myosins equivalent to human familial hypertrophic cardiomyopathy.
abstract: Familial hypertrophic cardiomyopathy (FHC) is caused by missence mutations in beta-myosin heavy chain or other various sarcomeric proteins. To elucidate the functional impact of FHC mutations in myosin heavy chain, we generated Dictyostelium discoideum myosin II mutants equivalent to human FHC mutations by site-directed mutagenesis, and characterized their molecular-basis motor function. The current mutants, i.e. R397Q, F506C, G575R, A699R, K703Q and K703W are equivalent to R403Q, F513C, G584R, G716R, R719Q and R719W FHC mutants respectively. We measured the molecular-basis force and the sliding velocity generated by these myosin mutants. The measurement revealed that the A699R, K703Q and K703W myosins exhibited the lowest level of force with their preserved actin-activated MgATPase activity. F506C mutant showed the least impairment of the motile and enzymatic activities. The motor function of R397Q and G575R myosins were classified as intermediate. These results suggest that ELC binding domain might be important for force production.
authors.affiliation: Second Department of Internal Medicine, University of Tokyo, Japan. fujitahtky@umin.ac.jp
authors.forename: H
authors.initials: H
authors.lastname: Fujita
authors.forename: S
authors.initials: S
authors.lastname: Sugiura
authors.forename: S
authors.initials: S
authors.lastname: Momomura
authors.forename: H
authors.initials: H
authors.lastname: Sugi
authors.forename: K
authors.initials: K
authors.lastname: Sutoh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002312:Cardiomyopathy, Hypertrophic / Q000235:genetics* / Q000378:metabolism; D004023:Dictyostelium; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009200:Myocardial Contraction / Q000235:genetics; D009218:Myosins / Q000235:genetics* / Q000378:metabolism; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D009218:Myosins