Actomyosin energy turnover declines while force remains constant during isometric muscle contraction.
Energy turnover was measured during isometric contractions of intact and Triton-permeabilized white fibres from dogfish (Scyliorhinus canicula) at 12 degrees C. Heat + work from actomyosin in intact fibres was determined from the dependence of heat + work output on filament overlap. Inorganic phosphate (Pi) release by permeabilized fibres was recorded using the fluorescent protein MDCC-PBP, N-(2-[1-maleimidyl]ethyl)-7-diethylamino-coumarin-3 carboxamide phosphate binding protein. The steady-state ADP release rate was measured using a linked enzyme assay. The rates decreased five-fold during contraction in both intact and permeabilized fibres. In intact fibres the rate of heat + work output by actomyosin decreased from 134 +/-s.e.m. 28 microW mg(-1) (n = 17) at 0.055 s to 42% of this value at 0.25 s, and to 20% at 3.5 s. The force remained constant between 0.25 and 3.5 s. Similarly in permeabilized fibres the Pi release rate decreased from 5.00 +/- 0.39 mmol l(-1) s(-1) at 0.055 s to 39% of this value at 0.25 s and to 19% at 0.5 s. The steady-state ADP release rate at 15 s was 21% of the Pi rate at 0.055 s. Using a single set of rate constants, the time courses of force, heat + work and Pi release were described by an actomyosin model that took account of the transition from the initial state (rest or rigor) to the contracting state, shortening and the consequent work against series elasticity, and reaction heats. The model suggests that increasing Pi concentration slows the cycle in intact fibres, and that changes in ATP and ADP slow the cycle in permeabilized fibres.
West
Timothy G
TG
Biological Structure & Function Section, Division of Biomedical Sciences, Sir Alexander Fleming Building, Imperial College London, South Kensington Campus, London SW7 2AZ.
Curtin
N A
NA
Ferenczi
Michael A
MA
He
Zhen-He
ZH
Sun
Yin-Biao
YB
Irving
Malcolm
M
Woledge
Roger C
RC
D000205:Actomyosin / Q000378:metabolism*; D000818:Animals; D004284:Dogfish; D004734:Energy Metabolism / Q000502:physiology*; D066298:In Vitro Techniques; D007537:Isometric Contraction / Q000502:physiology*; D018656:Muscle Fibers, Fast-Twitch / Q000378:metabolism*
D000205:Actomyosin