A new model for the surface arrangement of myosin molecules in tarantula thick filaments.
Three-dimensional reconstructions of the negatively stained thick filaments of tarantula muscle with a resolution of 50 A have previously suggested that the helical tracks of myosin heads are zigzagged, short diagonal ridges being connected by nearly axial links. However, surface views of lower contour levels reveal an additional J-shaped feature approximately the size and shape of a myosin head. We have modelled the surface array of myosin heads on the filaments using as a building block a model of a two-headed regulated myosin molecule in which the regulatory light chains of the two heads together form a compact head-tail junction. Four parameters defining the radius, orientation and rotation of each myosin molecule were varied. In addition, the heads were allowed independently to bend in a plane perpendicular to the coiled-coil tail at three sites, and to tilt with respect to the tail and to twist at one of these sites. After low-pass filtering, models were aligned with the reconstruction, scored by cross-correlation and refined by simulated annealing. Comparison of the geometry of the reconstruction and the distance between domains in the myosin molecule narrowed the choice of models to two main classes. A good match to the reconstruction was obtained with a model in which each ridge is formed from the motor domain of a head pointing to the bare zone together with the head-tail junction of a neighbouring molecule. The heads pointing to the Z-disc intermittently occupy the J-position. Each motor domain interacts with the essential and regulatory light chains of the neighbouring heads. A near-radial spoke in the reconstruction connecting the backbone to one end of the ridge can be identified as the start of the coiled-coil tail.
Offer
G
G
Division of Molecular and Cellular Biology, Department of Clinical Veterinary Science, University of Bristol, UK. g.w.offer@bristol.ac.uk
Knight
P J
PJ
Burgess
S A
SA
Alamo
L
L
Padrón
R
R
D008841:Actin Cytoskeleton / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D000818:Animals; D001665:Binding Sites; D003198:Computer Simulation; D008958:Models, Molecular; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D017434:Protein Structure, Tertiary; D013112:Spiders; D013329:Structure-Activity Relationship
D009218:Myosins