Myosin cross-bridge orientation in rigor and in the presence of nucleotide studied by electron spin resonance.
The tilt series electron spin resonance (ESR) spectrum from muscle fibers decorated with spin labeled myosin subfragment 1 (S1) was measured from fibers in rigor and in the presence of MgADP. ESR spectra were measured at low amplitude modulation of the static magnetic field to insure that a minimum of spectral lineshape distortion occurs. Ten tilt series ESR data sets were fitted simultaneously by the model-independent methodology described in the accompanying paper (Burghardt, T. P., and A. R. French, 1989. Biophys. J. 56:525-534). By this method the average and standard error in the mean of order parameters for the probe angular distribution were calculated for the two states of the fiber investigated. The average order parameters were used to reconstruct the probe angular distribution in two dimensions, one angular dimension corresponding to a polar angle measured relative to the fiber axis, and the other a torsional angular degree of freedom of the probe. We find that the probe angular distributions for the rigor and MgADP states of the fiber differ such that the rigor distribution is broader and shifted relative to the distribution in the presence of MgADP. The shape of the rigor distribution suggests the presence of two probe orientations, one similar to that in the presence of MgADP, and another at a different orientation. The shape of the distribution in the presence of MgADP suggests that the binding of the nucleotide to the rigor cross-bridge shifts the spin population into a more homogeneous one by causing a cross-bridge rotation.
Ajtai
K
K
Department of Biochemistry and Molecular Biology, Mayo Foundation, Rochester, Minnesota 55095.
French
A R
AR
Burghardt
T P
TP
D000244:Adenosine Diphosphate / Q000032:analysis; D000818:Animals; D004578:Electron Spin Resonance Spectroscopy; D008274:Magnesium; D009132:Muscles / Q000032:analysis; D009218:Myosins / Q000032:analysis*; D009711:Nucleotides / Q000032:analysis*; D011487:Protein Conformation; D011817:Rabbits
D009711:Nucleotides; D000244:Adenosine Diphosphate; D009218:Myosins; D008274:Magnesium