Regulation of force development studied by photolysis of caged ADP in rabbit skinned psoas fibers.
The present study examined the effects of Ca(2+) and strongly bound cross-bridges on tension development induced by changes in the concentration of MgADP. Addition of MgADP to the bath increased isometric tension over a wide range of [Ca(2+)] in skinned fibers from rabbit psoas muscle. Tension-pCa (pCa is -log [Ca(2+)]) relationships and stiffness measurements indicated that MgADP increased mean force per cross-bridge at maximal Ca(2+) and increased recruitment of cross-bridges at submaximal Ca(2+). Photolysis of caged ADP to cause a 0.5 mM MgADP jump initiated an increase in isometric tension under all conditions examined, even at pCa 6.4 where there was no active tension before ADP release. Tension increased monophasically with an observed rate constant, k(ADP), which was similar in rate and Ca(2+) sensitivity to the rate constant of tension re-development, k(tr), measured in the same fibers by a release-re-stretch protocol. The amplitude of the caged ADP tension transient had a bell-shaped dependence on Ca(2+), reaching a maximum at intermediate Ca(2+) (pCa 6). The role of strong binding cross-bridges in the ADP response was tested by treatment of fibers with a strong binding derivative of myosin subfragment 1 (NEM-S1). In the presence of NEM-S1, the rate and amplitude of the caged ADP response were no longer sensitive to variations in the level of activator Ca(2+). The results are consistent with a model in which ADP-bound cross-bridges cooperatively activate the thin filament regulatory system at submaximal Ca(2+). This cooperative interaction influences both the magnitude and kinetics of force generation in skeletal muscle.
Lu
Z
Z
Department of Physiology, University of Wisconsin, Madison, Wisconsin 53706, USA.
Swartz
D R
DR
Metzger
J M
JM
Moss
R L
RL
Walker
J W
JW
D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D008297:Male; D008954:Models, Biological; D009119:Muscle Contraction; D015879:Myosin Subfragments / Q000378:metabolism; D009578:Nitrobenzenes / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism; D010782:Photolysis; D018583:Pliability; D016658:Psoas Muscles / Q000378:metabolism*; D011817:Rabbits
D015879:Myosin Subfragments; D009578:Nitrobenzenes; C101520:P(2)-(1-(2-nitrophenyl)ethyl) ADP; D010710:Phosphates; D000244:Adenosine Diphosphate; D008274:Magnesium; D002118:Calcium