title: Distribution and structure-function relationship of myosin heavy chain isoforms in the adult mouse heart.
abstract: The two cardiac myosin heavy chain isoforms, alpha and beta, exhibit distinct functional characteristics and therefore may be distributed regionally within the heart to match the functional demands of a specific region. In adult mouse hearts, which predominantly express alpha-myosin heavy chain, we observed high concentrations of beta-myosin in distinct areas such as at the tip of papillary muscles and at the base close to the valvular annulus. In light of these distinct distribution patterns of the myosin isoforms, we subsequently explored the isoform-specific structure-function relationships of the myosins. The alpha- and beta-isoforms are 93% identical in amino acid sequence, but it remains unclear which of the nonidentical residues determines isoform functionality. We hypothesized that residues situated within or close to the actin-binding interface of the myosin head influence actin binding and thereby modulate actin-activated ATPase activity. A chimeric myosin was created containing beta-sequence from amino acid 417 to 682 within the alpha-backbone. In mice, approximately 70% of the endogenous cardiac protein was replaced with the chimeric myosin. Myofibrils containing chimeric myosin exhibited ATPase activities that were depressed to the levels observed in hearts expressing approximately 70% beta-myosin. In vitro motility assays showed that the actin filament sliding velocity generated by chimeric myosin was similar to that of alpha-myosin, almost twice the velocities observed with beta-myosin. These data indicate that this large domain sequence switch conferred beta-like actin-activated ATPase activities to the chimeric myosin, suggesting that this region is responsible for the distinct hydrolytic properties of these myosin isoforms.
authors.affiliation: Division of Molecular Cardiovascular Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio 45229, USA.
authors.forename: Maike
authors.initials: M
authors.lastname: Krenz
authors.forename: Sakthivel
authors.initials: S
authors.lastname: Sadayappan
authors.forename: Hanna E
authors.initials: HE
authors.lastname: Osinska
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Henry
authors.forename: Samantha
authors.initials: S
authors.lastname: Beck
authors.forename: David M
authors.initials: DM
authors.lastname: Warshaw
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Robbins
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D051379:Mice; D009206:Myocardium / Q000737:chemistry* / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D014018:Tissue Distribution; D024723:Ventricular Myosins
chemical_list: D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D000251:Adenosine Triphosphatases; D024723:Ventricular Myosins; D018995:Myosin Heavy Chains