The active cross-bridge motions of isolated thick filaments from myosin-regulated muscles detected by quasi-elastic light scattering.
Intensity fluctuation spectroscopy has been used successfully as a probe that can detect an increase in high-frequency internal motions of isolated thick filaments of Limulus muscle upon the addition of calcium ions. We have attributed such motions to cross-bridge motion instead of to an increase in the flexibility of the filament backbone. Here we show that after cleavage of the S-1 and then the S-2 moieties with papain, cross-linking the myosin heads to the filament backbone, or heat denaturation (42 degrees C, 10 min), the increase in the high frequency internal motions in the thick filaments no longer occurs. Congo Red, which has been shown to induce shortening of isolated myofibrils, also increases the high-frequency motions of the isolated filaments. Furthermore, the increase is suppressed by treating the filaments with a myosin ATPase inhibitor such as vanadate ions (10 mM) or by replacing ATP with either an equimolar CrADP or the nonhydrolyzable ATP analogue beta, gamma-imido-adenine-5'-triphosphate (AMP-PNP). Calcium ions have a similar effect on isolated thick filaments from scallop muscle, where the myosin is known to be regulatory. Calcium ions have no such effect on thick filaments isolated from frog muscle, which is believed not to be regulated by calcium binding to myosin. These results confirm our earlier supposition that the additional high frequency internal motions of the thick filaments isolated from striated muscle of Limulus are related to the energy dependent, active cross-bridge motions.
Fan
S F
SF
Dewey
M M
MM
Colflesh
D
D
Gaylinn
B
B
Greguski
R A
RA
Chu
B
B
D000251:Adenosine Triphosphatases / Q000502:physiology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000266:Adenylyl Imidodiphosphate / Q000494:pharmacology; D000818:Animals; D003224:Congo Red / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D004548:Elasticity; D006737:Horseshoe Crabs; D008433:Mathematics; D008954:Models, Biological; D008974:Mollusca; D009132:Muscles / Q000502:physiology*; D009218:Myosins / Q000502:physiology*; D011894:Rana pipiens; D012542:Scattering, Radiation
D000266:Adenylyl Imidodiphosphate; D003224:Congo Red; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D009218:Myosins