Monitoring the Myosin Crossbridge Cycle in Contracting Muscle: Steps towards ‘Muscle the Movie’.

Felicity Eakins, Carlo Knupp, John M Squire

Research output: Contribution to journalArticle (Academic Journal)

2 Citations (Scopus)
4 Downloads (Pure)

Abstract

Some vertebrate muscles (e.g. those in bony fsh) have a simple lattice A-band which is so well ordered that low-angle X-ray difraction patterns are sampled in a simple way amenable to crystallographic techniques. Time-resolved X-ray diffraction through the contractile cycle should provide a movie of the molecular movements involved in muscle contraction. Generation of ‘Muscle—The Movie’ was suggested in the 1990s and since then eforts have been made to work out how to achieve it. Here we discuss how a movie can be generated, we discuss the problems and opportunities, and present some new observations. Low angle X-ray difraction patterns from bony fsh muscles show myosin layer lines that are well sampled on row-lines expected from the simple hexagonal A-band lattice. The 1st, 2nd and 3rd myosin layer lines at d-spacings of around 42.9 nm, 21.5 nm and 14.3 nm respectively, get weaker in patterns from active muscle, but there is a well-sampled intensity remnant along the layer lines. We show here that the pattern from the tetanus plateau is not a residual resting pattern from fbres that have not been fully activated, but is a diferent well-sampled pattern showing the presence of a second, myosin-centred, arrangement of crossbridges within the active crossbridge population. We also show that the meridional M3 peak from active muscle has two components of diferent radial widths consistent with (i) active myosin-centred (probably weak-binding) heads giving a narrow peak and (ii) heads on actin in strong states giving a broad peak.
Original languageEnglish
Pages (from-to)77 - 91
Number of pages15
JournalJournal of Muscle Research and Cell Motility
Volume40
Early online date20 Jul 2019
DOIs
Publication statusE-pub ahead of print - 20 Jul 2019

Keywords

  • Time-resolved X-ray difraction from muscle
  • Myosin head organisation
  • Muscle lattice disorder
  • Weakbinding myosin head state
  • Interacting heads motif
  • Muscle M3 meridional peak

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