Motion of subfragment-1 in myosin and its supramolecular complexes: saturation transfer electron paramagnetic resonance.
Molecular dynamics in spin-labeled muscle proteins was studied with a recently developed electron paramagnetic resonance (EPR) technique, saturation transfer spectroscopy, which is uniquely sensitive to rotational motion in the range of 10(-7)-10(-3) sec. Rotational correlation time (tau2) were determined for a spin label analog of iodoacetamide bound to the subfragment-1 (S-1) region of myosin under a variety of conditions likely to shed light on the molecular mechanism of muscle contraction. Results show that (a) the spin labels are rigidly bound to the isolated S-1 (tau2 = 2 x 10(-7) sec) and can be used to estimate values of tau2 for the S-1 region of the myosin molecule; (b) in solutions of intact myosin, S-1 has considerable mobility relative to the rest of the myosin molecule, the value of tau2 for the S-1 segment of myosin being less than twice that for isolated S-1, while the molecular weights differ by a factor of 4 to 5; (c) in myosin filaments, tau2 increases by a factor of only about 10, suggesting motion of the S-1 regions independent of the backbone of the myosin filament, but slower than that in a single molecule; (d) addition of F-actin to solutions of myosin or S-1 increases tau2 by a factor of nearly 10(3), indicating strong immobilization of S-1 upon binding to actin. Saturation transfer spectroscopy promises to provide an extremely useful tool for the study of the motions of the crossbridges and thin filaments in reconstituted systems and in glycerinated muscle fibers.
D001665:Binding Sites; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D005033:Ethylmaleimide; D007460:Iodoacetamide; D046911:Macromolecular Substances; D009218:Myosins; D010446:Peptide Fragments; D011485:Protein Binding; D011487:Protein Conformation; D013113:Spin Labels; D013438:Sulfhydryl Compounds; D013997:Time Factors; D014357:Trypsin
D046911:Macromolecular Substances; D010446:Peptide Fragments; D013113:Spin Labels; D013438:Sulfhydryl Compounds; D014357:Trypsin; D009218:Myosins; D005033:Ethylmaleimide; D007460:Iodoacetamide