CHARACTERIZATIONS OF CROSS-BRIDGES IN THE PRESENCE OF SATURATING CONCENTRATIONS OF MGAMP-PNP IN RABBIT PERMEABILIZED PSOAS MUSCLE

Several earlier studies have led to different conclusions about the co mplex of myosin with MgAMP-PNP. It has been suggested that subfragment 1 of myosin (S1)-MSAMP-PNP forms an S1-MgADP-like state, an intermedi ate between the myosin S1-MgATP and myosin S1-MgADP states or a mixtur e of cross-bridge states. We suggest that the different states observe d result from the failure to saturate S1 with MgAMP-PNP. At saturating MgAMP-PNP, the interaction of myosin S1 with actin is very similar to that which occurs in the presence of MgATP. 1) At 1 degrees C and 170 mM ionic strength the equatorial x-ray diffraction intensity ratio I- 11/I-10 decreased with an increasing MgAMP-PNP concentration and level ed off by similar to 20 mM MgAMP-PNP. The resulting ratio was the same for MgATP-relaxed fibers. 2) The two dimensional x-ray diffraction pa tterns from MgATP-relaxed and MgAMP-PNP-relaxed bundles are similar. 3 ) The affinity of S1-MgAMP-PNP for the actin-tropomyosin-troponin comp lex in solution in the absence of free calcium is comparable with that of S1-MgATP. 4) In the presence of calcium, I-11/I-10 decreased towar d the relaxed value with increasing MgAMP-PNP, signifying that the aff inity between cross-bridge and actin is weakened by MgAMP-PNP. 5) The degree to which the equatorial intensity ratio decreases as the ionic strength increases is similar in MgAMP-PNP and MgATP. Therefore, resul ts from both fiber and solution studies suggest that MgAMP-PNP acts as a non hydrolyzable MgATP analogue for myosin.