A1603P and K1617del, Mutations in β-Cardiac Myosin Heavy Chain that Cause Laing Early-Onset Distal Myopathy, Affect Secondary Structure and Filament Formation In Vitro and In Vivo. Issue 10 (11th May 2018)

• Record Type: Journal Article
• Authors: Parker, Francine; Batchelor, Matthew; Wolny, Marcin; Hughes, Ruth; Knight, Peter J.; Peckham, Michelle
• Is Part Of: Journal of molecular biology. Volume 430:Issue 10(2018)
• Page Start: 1459
• Page End: 1478
• Publication Date: 2018-05-11
• Subjects: myosin -- myopathy -- muscle -- coiled coil -- mutations; β-MHC β-cardiac myosin heavy chain -- MPD-1 Laing early-onset distal myopathy -- GST glutathione S-transferase -- LMM light meromyosin -- EM electron microscopy -- CD circular dichroism -- WT wild-type -- MD molecular dynamics -- RMSF root mean square fluctuation -- MRE mean residue ellipticity; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2018.04.006
• Languages: English
• ISSNs: 0022-2836

Abstract

Abstract: Over 20 mutations in β-cardiac myosin heavy chain (β-MHC), expressed in cardiac and slow muscle fibers, cause Laing early-onset distal myopathy (MPD-1), a skeletal muscle myopathy. Most of these mutations are in the coiled-coil tail and commonly involve a mutation to a proline or a single-residue deletion, both of which are predicted to strongly affect the secondary structure of the coiled coil. To test this, we characterized the effects of two MPD-1 causing mutations: A1603P and K1617del in vitro and in cells. Both mutations affected secondary structure, decreasing the helical content of 15 heptad and light meromyosin constructs. Both mutations also severely disrupted the ability of glutathione S -transferase–light meromyosin fusion proteins to form minifilaments in vitro, as demonstrated by negative stain electron microscopy. Mutant eGFP-tagged β-MHC accumulated abnormally into the M-line of sarcomeres in cultured skeletal muscle myotubes. Incorporation of eGFP-tagged β-MHC into sarcomeres in adult rat cardiomyocytes was reduced. Molecular dynamics simulations using a composite structure of part of the coiled coil demonstrated that both mutations affected the structure, with the mutation to proline (A1603P) having a smaller effect compared to K1617del. Taken together, it seems likely that the MPD-1 mutations destabilize the coiled coil, resulting in aberrant myosin packing in thick filaments in muscle sarcomeres, providing a potential mechanism for the disease. Graphical abstract: Highlights: It is unclear how mutations in the coiled coil of β-myosin heavy chain cause distal myopathy. A1603P and K1617del mutations reduce helicity and affect filament formation in vitro . eGFP-tagged β-myosin heavy chain abnormally accumulates at the M-line of sarcomeres in skeletal myotubes. Molecular dynamics simulations provide a molecular understanding for these experiments. Effects on structure and packing into the thick filament provide a molecular basis for the disease.