Changes in myosin S1 orientation and force induced by a temperature increase.
Force generation in myosin-based motile systems is thought to result from an angular displacement of the myosin subfragment 1 (S1) tail domain with respect to the actin filament axis. In muscle, raised temperature increases the force generated by S1, implying a greater change in tail domain angular displacement. We used time-resolved x-ray diffraction to investigate the structural corollary of this force increase by measuring M3 meridional reflection intensity during sinusoidal length oscillations. This technique allows definition of S1 orientation with respect to the myofilament axis. M3 intensity changes were approximately sinusoid at low temperatures but became increasingly distorted as temperature was elevated, with the formation of a double intensity peak at maximum shortening. This increased distortion could be accounted for by assuming a shift in orientation of the tail domain of actin-bound S1 toward the orientation at which M3 intensity is maximal, which is consistent with a tail domain rotation model of force generation in which the tail approaches a more perpendicular projection from the thin filament axis at higher temperatures. In power stroke simulations, the angle between S1 tail mean position during oscillations and the position at maximum intensity decreased by 4.7 degrees, corresponding to a mean tail displacement toward the perpendicular of 0.73 nm for a temperature-induced force increase of 0.28 P(0) from 4 to 22 degrees C. Our findings suggest that at least 62% of crossbridge compliance is associated with the tail domain.
Griffiths
Peter J
PJ
Dipartimento di Scienze Fisiologiche, Università degli Studi di Firenze, Viale G.B. Morgagni 63, I-50132 Florence, Italy. pjg@physiol.ox.ac.uk
Bagni
Maria A
MA
Colombini
Barbara
B
Amenitsch
Heinz
H
Bernstorff
Sigrid
S
Ashley
Christopher C
CC
Cecchi
Giovanni
G
Ameritsch
Heinz
H
D000199:Actins / Q000737:chemistry; D000465:Algorithms; D000818:Animals; D007700:Kinetics; D007834:Lasers; D009132:Muscles / Q000378:metabolism; D015879:Myosin Subfragments / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D011896:Rana temporaria; D013696:Temperature; D013816:Thermodynamics; D014961:X-Ray Diffraction; D014965:X-Rays
D000199:Actins; D015879:Myosin Subfragments