title: Orientation of the N-terminal lobe of the myosin regulatory light chain in skeletal muscle fibers.
abstract: The orientation of the N-terminal lobe of the myosin regulatory light chain (RLC) in demembranated fibers of rabbit psoas muscle was determined by polarized fluorescence. The native RLC was replaced by a smooth muscle RLC with a bifunctional rhodamine probe attached to its A, B, C, or D helix. Fiber fluorescence data were interpreted using the crystal structure of the head domain of chicken skeletal myosin in the nucleotide-free state. The peak angle between the lever axis of the myosin head and the fiber or actin filament axis was 100-110° in relaxation, isometric contraction, and rigor. In each state the hook helix was at an angle of ∼40° to the lever/filament plane. The in situ orientation of the RLC D and E helices, and by implication of its N- and C-lobes, was similar in smooth and skeletal RLC isoforms. The angle between these two RLC lobes in rigor fibers was different from that in the crystal structure. These results extend previous crystallographic evidence for bending between the two lobes of the RLC to actin-attached myosin heads in muscle fibers, and suggest that such bending may have functional significance in contraction and regulation of vertebrate striated muscle.
authors.affiliation: Medical Research Council, National Institute for Medical Research, London, United Kingdom.
authors.forename: Daniela
authors.initials: D
authors.lastname: Romano
authors.forename: Birgit D
authors.initials: BD
authors.lastname: Brandmeier
authors.forename: Yin-Biao
authors.initials: YB
authors.lastname: Sun
authors.forename: David R
authors.initials: DR
authors.lastname: Trentham
authors.forename: Malcolm
authors.initials: M
authors.lastname: Irving
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002645:Chickens; D002942:Circular Dichroism; D018485:Muscle Fibers, Skeletal / Q000378:metabolism*; D009130:Muscle, Smooth / Q000378:metabolism; D018994:Myosin Light Chains / Q000737:chemistry* / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D017433:Protein Structure, Secondary; D011817:Rabbits; D012235:Rhodamines / Q000378:metabolism; D013237:Stereoisomerism
chemical_list: D018994:Myosin Light Chains; D020033:Protein Isoforms; D012235:Rhodamines; D000251:Adenosine Triphosphatases