Myosin filament assembly requires a cluster of four positive residues located in the rod domain.
Myosin has an intrinsic ability to organize into ordered thick filaments that mediate muscle contraction. Here, we use surface plasmon resonance and light scattering analysis to further characterize the molecular determinants that guide myosin filament assembly. Both assays identify a cluster of lysine and arginine residues as important for myosin polymerization in vitro. Moreover, in cardiomyocytes, replacement of these charged residues by alanine severely affects the incorporation of myosin into the distal ends of the sarcomere. Our findings show that a novel assembly element with a distinct charge profile is present at the C-terminus of sarcomeric myosins.
Department of Molecular, Cellular, and Developmental Biology and BioFrontiers Institute, University of Colorado, Boulder, CO 80303, United States.
Robert C
RC
Thompson
Massimo
M
Buvoli
Ada
A
Buvoli
Leslie A
LA
Leinwand
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D004563:Electrochemistry; D008027:Light; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000737:chemistry / Q000378:metabolism; D015879:Myosin Subfragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012542:Scattering, Radiation; D020349:Surface Plasmon Resonance; D014162:Transfection; D024723:Ventricular Myosins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D050505:Mutant Proteins; D015879:Myosin Subfragments; D011993:Recombinant Fusion Proteins; D024723:Ventricular Myosins