Steady-state kinetics of MgATP splitting by native myosin RLC-free subfragment 1.

MgATP positively regulates the dimerisation reaction, resulting in an increase in the rate of MgATP splitting with increasing MgATP concentration. We investigated the stoichiometry of this dimerisation reaction and found that each subunit in the dimer bound one molecule of MgATP at the dimerisation site. We studied changes with temperature in the MgATPase activity of S1 in the dimeric form for temperatures of 18-25 degrees C. Between 18.0 and 21.2 degrees C, kcat increased steadily with temperature. Between 21.2 and 21.8 degrees C, there was a large decrease in kcat. A strong increase in kcat occurred at temperatures above 21.8 degrees C, corresponding to a new reversible conformation of S1, unable to dimerise. The steep decrease in kcat between 21.2 and 21.8 degrees C is due to a temperature transition in the monomer-dimer equilibrium.