Orientation of myosin binding protein C in the cardiac muscle sarcomere determined by domain-specific immuno-EM
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UMass Chan AffiliationsDepartment of Cell and Developmental Biology
Document TypeJournal Article
Image Processing, Computer-Assisted
cardiac muscle contraction
cardiac muscle disease
cardiac muscle regulation
cardiac muscle structure
Cell and Developmental Biology
Cellular and Molecular Physiology
MetadataShow full item record
AbstractMyosin binding protein C is a thick filament protein of vertebrate striated muscle. The cardiac isoform [cardiac myosin binding protein C (cMyBP-C)] is essential for normal cardiac function, and mutations in cMyBP-C cause cardiac muscle disease. The rod-shaped molecule is composed primarily of 11 immunoglobulin- or fibronectin-like domains and is located at nine sites, 43nm apart, in each half of the A-band. To understand how cMyBP-C functions, it is important to know its structural organization in the sarcomere, as this will affect its ability to interact with other sarcomeric proteins. Several models, in which cMyBP-C wraps around, extends radially from, or runs axially along the thick filament, have been proposed. Our goal was to define cMyBP-C orientation by determining the relative axial positions of different cMyBP-C domains. Immuno-electron microscopy was performed using mouse cardiac myofibrils labeled with antibodies specific to the N- and C-terminal domains and to the middle of cMyBP-C. Antibodies to all regions of the molecule, except the C-terminus, labeled at the same nine axial positions in each half A-band, consistent with a circumferential and/or radial rather than an axial orientation of the bulk of the molecule. The C-terminal antibody stripes were slightly displaced axially, demonstrating an axial orientation of the C-terminal three domains, with the C-terminus closer to the M-line. These results, combined with previous studies, suggest that the C-terminal domains of cMyBP-C run along the thick filament surface, while the N-terminus extends toward neighboring thin filaments. This organization provides a structural framework for understanding cMyBP-C's modulation of cardiac muscle contraction.
SourceJ Mol Biol. 2015 Jan 30;427(2):274-86. doi: 10.1016/j.jmb.2014.10.023. Link to article on publisher's site.
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/26454
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