Characterization of the cross-bridge force-generating step using inorganic phosphate and BDM in myofibrils from rabbit skeletal muscles.
The inhibitory effects of inorganic phosphate (P(i)) on isometric force in striated muscle suggest that in the ATPase reaction P(i) release is coupled to force generation. Whether P(i) release and the power stroke are synchronous events or force is generated by an isomerization of the quaternary complex of actomyosin and ATPase products (AM.ADP.P(i)) prior to the following release of P(i) is still controversial. Examination of the dependence of isometric force on [P(i)] in rabbit fast (psoas; 5-15 degrees C) and slow (soleus; 15-20 degrees C) myofibrils was used to test the two-step hypothesis of force generation and P(i) release. Hyperbolic fits of force-[P(i)] relations obtained in fast and slow myofibrils at 15 degrees C produced an apparent asymptote as [P(i)]-->infinity of 0.07 and 0.44 maximal isometric force (i.e. force in the absence of P(i)) in psoas and soleus myofibrils, respectively, with an apparent K(d) of 4.3 mM in both. In each muscle type, the force-[P(i)] relation was independent of temperature. However, 2,3-butanedione 2-monoxime (BDM) decreased the apparent asymptote of force in both muscle types, as expected from its inhibition of the force-generating isomerization. These data lend strong support to models of cross-bridge action in which force is produced by an isomerization of the AM.ADP.P(i) complex immediately preceding the P(i) release step.
Tesi
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Dipartimento di Scienze Fisiologiche, Università degli Studi di Firenze, Viale GB, Morgagni 63, I-50134 Firenze, Italy. chiara.tesi@unifi.it
Colomo
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Piroddi
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N
Poggesi
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C
D000244:Adenosine Diphosphate / Q000502:physiology; D000465:Algorithms; D000818:Animals; D003931:Diacetyl / Q000031:analogs & derivatives* / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D066298:In Vitro Techniques; D007536:Isomerism; D007537:Isometric Contraction / Q000187:drug effects / Q000502:physiology; D018656:Muscle Fibers, Fast-Twitch / Q000187:drug effects / Q000502:physiology; D018657:Muscle Fibers, Slow-Twitch / Q000187:drug effects / Q000502:physiology; D018482:Muscle, Skeletal / Q000187:drug effects / Q000502:physiology*; D009210:Myofibrils / Q000187:drug effects / Q000502:physiology*; D010710:Phosphates / Q000494:pharmacology*; D011817:Rabbits; D013696:Temperature
D004791:Enzyme Inhibitors; D010710:Phosphates; C004717:diacetylmonoxime; D000244:Adenosine Diphosphate; D003931:Diacetyl