The myosin II coiled-coil domain atomic structure in its native environment

Abstract

Significance Myosin II is the molecule that produces force in muscle contraction. Unlike the myosin head, its molecular motor, no atomic resolution structure of the ∼1000-residue–long α-helical coiled-coil tail has been reported. Here, we describe the cryo-EM atomic structure of the myosin tail within a native muscle thick filament. Three differences with crystal structures of myosin tail segments were found. The myosin head arrangement apparently alters the beginning of the tail. Striated muscle myosins have four skip residues, amino acids inserted to improve the alignment of charged residue clusters. Skips 1 and 3 agree with the crystal structures. Skip 2, which is a novel structure, and Skip 4 do not. Functional consequences are suggested by the myosin tail packing. The atomic structure of the complete myosin tail within thick filaments isolated from Lethocerus indicus flight muscle is described and compared to crystal structures of recombinant, human cardiac myosin tail segments. Overall, the agreement is good with three exceptions: the proximal S2, in which the filament has heads attached but the crystal structure doesn’t, and skip regions 2 and 4. At the head–tail junction, the tail α-helices are asymmetrically structured encompassing well-defined unfolding of 12 residues for one myosin tail, ∼4 residues of the other, and different degrees of α-helix unwinding for both tail α-helices, thereby providing an atomic resolution description of coiled-coil “uncoiling” at the head–tail junction. Asymmetry is observed in the nonhelical C termini; one C-terminal segment is intercalated between ribbons of myosin tails, the other apparently terminating at Skip 4 of another myosin tail. Between skip residues, crystal and filament structures agree well. Skips 1 and 3 also agree well and show the expected α-helix unwinding and coiled-coil untwisting in response to skip residue insertion. Skips 2 and 4 are different. Skip 2 is accommodated in an unusual manner through an increase in α-helix radius and corresponding reduction in rise/residue. Skip 4 remains helical in one chain, with the other chain unfolded, apparently influenced by the acidic myosin C terminus. The atomic model may shed some light on thick filament mechanosensing and is a step in understanding the complex roles that thick filaments of all species undergo during muscle contraction.