Ligand-induced myosin subfragment 1 global conformational change.
The effects of selected ligands on the structure of myosin subfragment 1 (S1) were compared by using transient electrical birefringence techniques. With pairs of dilute solutions of S1 at 3.5 degrees C in low ionic strength (mu = 0.020 M) buffers that had matched electrical impedances, S1 with Mg2+, MgADP, or MgADP.Vi bound was subjected to 6-7-microseconds external electrical fields in the Kerr law range. Specific Kerr constants and the rates of rotational Brownian motion after the electric field was removed were measured. Neither Mg2+ nor MgADP had a measurable effect on either observable, but when orthovanadate (Vi) bound S1.MgADP it decreased the rotational correlation coefficient from 267 +/- 6 to 244 +/- 10 ns. Parallel measurements of MgATPase activity indicated that S1.MgADP.Vi was greater than 95% inhibited. These results confirm the conclusion of Aguirre et al. [(1989) Biochemistry 28, 799] that Vi binding to S1.MgADP increases its rate of rotational Brownian motion and provide data that are more quantitatively correlated with S1 structure. The Vi-induced change in the rotational correlation coefficient is consistent with S1 becoming more flexible or more compact when Vi binds. Assuming that S1.MgADP.Vi is an analogue for S1.MgADP.Pi, the structural changes observed for S1-ligand complexes in solution are discussed in relation to possible structural changes of intermediates on the kinetic pathway of ATPase hydrolysis. A new model of force generation by S1 in muscle is hypothesized.
Highsmith
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Department of Biochemistry, University of the Pacific, School of Dentistry, San Francisco, California 94115.
Eden
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D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001718:Birefringence; D004058:Diffusion; D004563:Electrochemistry; D006868:Hydrolysis; D007700:Kinetics; D008024:Ligands; D008274:Magnesium / Q000378:metabolism; D015879:Myosin Subfragments / Q000378:metabolism*; D011487:Protein Conformation; D011817:Rabbits; D012399:Rotation; D013329:Structure-Activity Relationship
D008024:Ligands; D015879:Myosin Subfragments; D000255:Adenosine Triphosphate; D008274:Magnesium