title: Functional analysis of myosin missense mutations in familial hypertrophic cardiomyopathy.
abstract: To analyze potential functional consequences of myosin heavy chain (MHC) mutations identified in patients with familial hypertrophic cardiomyopathy (FHC), we have assessed the stability of the mutant MHCs and their ability to form thick filaments. Constructs encoding wild-type rat alpha MHC and seven corresponding FHC missense mutants were transfected into COS cells. Immunoblot analysis suggested that FHC mutations do not grossly alter protein stability. Wild-type alpha MHC transfected into COS cells forms structures previously shown to be arrays of thick filaments, which also resemble myosin structures observed early in differentiation of muscle cells. Surprisingly, up to 29% of COS cells transfected with the FHC mutants failed to form filamentous structures. To assess whether this phenotype was specific for the FHC mutants and not generalizable to any myosin mutation, COS cells were transfected with a construct encoding an MHC with a 168-amino acid deletion of the hinge/rod region. This deletion construct formed filamentous structures with the same frequency as wild-type MHC. Biochemical analysis of one FHC mutant (Arg-249-->Gln) demonstrates that the structures formed by the mutant are solubilized at a lower ionic strength than those formed by wild-type MHC. We conclude that although the FHC mutant MHC is not labile, its assembly properties may be impaired.
authors.affiliation: Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: A J
authors.initials: AJ
authors.lastname: Straceski
authors.forename: A
authors.initials: A
authors.lastname: Geisterfer-Lowrance
authors.forename: C E
authors.initials: CE
authors.lastname: Seidman
authors.forename: J G
authors.initials: JG
authors.lastname: Seidman
authors.forename: L A
authors.initials: LA
authors.lastname: Leinwand
mesh_terms: D000818:Animals; D002312:Cardiomyopathy, Hypertrophic / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002460:Cell Line; D002522:Chlorocebus aethiops; D006801:Humans; D008854:Microscopy, Electron; D009218:Myosins / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D010641:Phenotype; D017354:Point Mutation; D051381:Rats; D012995:Solubility; D014162:Transfection
chemical_list: D009218:Myosins