Correlation between mechanical and enzymatic events in contracting skeletal muscle fiber.
The conventional hypothesis of muscle contraction postulates that the interaction between actin and myosin involves tight coupling between the power stroke and hydrolysis of ATP. However, some in vitro experiments suggested that hydrolysis of a single molecule of ATP caused multiple mechanical cycles. To test whether the tight coupling is present in contracting muscle, we simultaneously followed mechanical and enzymatic events in a small population of cross-bridges of glycerinated rabbit psoas fibers. Such small population behaves as a single cross-bridge when muscle contraction is initiated by a sudden release of caged ATP. Mechanical events were measured by changes of orientation of probes bound to the regulatory domain of myosin. Enzymatic events were simultaneously measured from the same cross-bridge population by the release of fluorescent ADP from the active site. If the conventional view were true, ADP desorption would occur simultaneously with dissociation of cross-bridges from thin filaments and would be followed by cross-bridge rebinding to thin filaments. Such sequence of events was indeed observed in contracting muscle fibers, suggesting that mechanical and enzymatic events are tightly coupled in vivo.
Department of Molecular Biology and Immunology, University of North Texas, 3500 Camp Bowie Boulevard, Fort Worth, Texas 7610, USA.
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Shepard
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Borejdo
D000244:Adenosine Diphosphate / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000737:chemistry; D000818:Animals; D001696:Biomechanical Phenomena; D005454:Fluorescence Polarization; D007700:Kinetics; D009119:Muscle Contraction / Q000502:physiology*; D018485:Muscle Fibers, Skeletal / Q000201:enzymology* / Q000502:physiology; D018994:Myosin Light Chains / Q000737:chemistry; D009218:Myosins / Q000737:chemistry* / Q000502:physiology; D016658:Psoas Muscles / Q000201:enzymology* / Q000502:physiology; D011817:Rabbits; D012235:Rhodamines / Q000737:chemistry
D018994:Myosin Light Chains; D012235:Rhodamines; D000244:Adenosine Diphosphate; C029273:P(3)-1-(2-nitro)phenylethyladenosine 5'-triphosphate; C034996:tetramethylrhodamine iodoacetamide; D000255:Adenosine Triphosphate; D009218:Myosins