Angle of active site of myosin heads in contracting muscle during sudden length changes.
The change in orientation of myosin crossbridges in contracting muscle during sudden length changes was examined by fluorescence polarization. This study used a fluorescent ATP analogue, 1,N6-etheno-2-aza-ATP(epsilon-2-aza-ATP) as a probe. Its fluorescence is considerably enhanced upon binding with myosin and is dependent on the chemical state of the myosin-nucleotide complex in muscle. The results showed that nucleotides bound to crossbridges in the intermediate attached state (presumably AM-epsilon-2-aza-ADP-Pi) during isometric contraction are highly oriented at the same angle as that of AM in rigor with bound epsilon-2-aza-ADP. Furthermore the orientation of nucleotides bound to crossbridges in the attached state is not altered during sudden changes in length of isometrically contracting muscle. The results of this time-resolved measurement support the conclusion obtained from a previous steady-state experiment that change in axial orientation of the active site of the myosin head is not involved in force generation.
D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D004985:Ethenoadenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D007537:Isometric Contraction / Q000187:drug effects; D007700:Kinetics; D009119:Muscle Contraction / Q000187:drug effects; D009126:Muscle Relaxation / Q000187:drug effects; D009132:Muscles / Q000502:physiology*; D009218:Myosins / Q000378:metabolism*; D011817:Rabbits; D013050:Spectrometry, Fluorescence
D004985:Ethenoadenosine Triphosphate; C012758:aza-epsilon-ATP; D009218:Myosins; D002118:Calcium