A birefringence study of changes in myosin orientation during relaxation of skinned muscle fibers induced by photolytic ATP release.
The birefringence of isolated skinned fibers from rabbit psoas muscle was measured continuously during relaxation from rigor produced by photolysis of caged ATP at sarcomere length 2.8-2.9 microns, ionic strength 0.1 M, 15 degrees C. Birefringence, the difference in refractive index between light components polarized parallel and perpendicular to the fiber axis, depends on the average degree of alignment of the myosin head domain with the fiber axis. After ATP release birefringence increased by 5.8 +/- 0.7% (mean +/- SE, n = 6) with two temporal components. A small fast component had an amplitude of 0.9 +/- 0.2% and rate constant of 63 s-1. By the completion of this component, the instantaneous stiffness had decreased to about half the rigor value, and the force response to a step stretch showed a rapid (approximately 1000 s-1) recovery phase. Subsequently a large slow birefringence component with rate constant 5.1 s-1 accompanied isometric force relaxation. Inorganic phosphate (10 mM) did not affect the fast birefringence component but accelerated the slow component and force relaxation. The fast birefringence component was probably caused by formation of myosin.ATP or myosin.ADP.Pi states that are weakly bound to actin. The average myosin head orientation at the end of this component is slightly more parallel to the fiber axis than in rigor.
Peckham
M
M
Randall Institute, King's College London.
Ferenczi
M A
MA
Irving
M
M
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001696:Biomechanical Phenomena; D055592:Biophysical Phenomena; D001703:Biophysics; D001718:Birefringence; D004548:Elasticity; D066298:In Vitro Techniques; D018485:Muscle Fibers, Skeletal / Q000737:chemistry* / Q000502:physiology; D009126:Muscle Relaxation / Q000187:drug effects / Q000502:physiology*; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D010710:Phosphates / Q000494:pharmacology; D010782:Photolysis; D016658:Psoas Muscles / Q000737:chemistry / Q000502:physiology; D011817:Rabbits; D012518:Sarcomeres / Q000737:chemistry / Q000502:physiology
D010710:Phosphates; C029273:P(3)-1-(2-nitro)phenylethyladenosine 5'-triphosphate; D000255:Adenosine Triphosphate; D009218:Myosins