X-Ray solution scattering of squid heavy meromyosin: Strengthening the evidence for an ancient compact off state

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The overall conformations of regulated myosins or heavy meromyosins from chicken/turkey, scallop, tarantula, limulus, and scorpion sources have been studied by a number of techniques, including electron microscopy, sedimentation, and pulsed electron paramagnetic resonance. These studies have indicated that the binding of regulatory ions changes the conformation of the molecule from a compact shape found in the "off" state of the muscle to extended relationships between the tail and independently mobile heads that predominate in the "on" state. Here we strengthen the argument for the generality of this conformational change by using small angle X-ray scattering on heavy meromyosin from squid. Small angle X-ray scattering allows the protein to be visualized in solution under mild and relatively physiological conditions, and squid differs from the other species studied by at least 500 million years of evolution. Analysis of the data indicates that upon addition of Ca2+ the radius of gyration increases. Differences in the squid "on" and "off" states are clearly distinguishable as bimodal and unimodal pair distance distribution functions respectively. These observations are consistent with a Ca2+-free squid heavy meromyosin that is compact, but which becomes extended when Ca2+ is bound. Further, the scattering profile derived from the current model of tarantula heavy meromyosin in the "off" state is in excellent agreement with the measured "off" state scattering profile for squid heavy meromyosin. The previous and current studies together provide significant evidence that regulated myosin's compact off-state conformation is an ancient trait, inherited from a common ancestor during divergent evolution. © 2013 Gillilan et al.




Gillilan, R. E., Senthil Kumar, V. S., O’Neall-Hennessey, E., Cohen, C., & Brown, J. H. (2013). X-Ray solution scattering of squid heavy meromyosin: Strengthening the evidence for an ancient compact off state. PLoS ONE, 8(12). https://doi.org/10.1371/journal.pone.0081994

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