The polymerization of actin: Extent of polymerization under pressure, volume change of polymerization, and relaxation after temperature jumps

The protein actin can polymerize from monomeric globular G-actin to polymeric filamentary F-actin, under the regulation of thermodynamic variables such as temperature, pressure, and compositions of G-actin and salts. We present here new measurements of the extent of polymerization (phi) of actin under pressure (P), for rabbit skeletal muscle actin in H2O buffer in the presence of adenosine triposphate and calcium ions and at low (5-15 mM) KCl concentrations. We measured phi using pyrene-labeled actin, as a function of time (t) and temperature (T), for samples of fixed concentrations of initial G-actin and KCl and at fixed pressure. The phi(T,P) measurements at equilibrium have the same form as reported previously at 1 atm: low levels of polymerization at low temperatures, representing dimerization of the actin; an increase in phi at the polymerization temperature (T-p); a maximum in phi(T) above T-p with a decrease in phi(T) beyond the maximum, indicating a depolymerization at higher T. From phi(T,P) at temperatures below T-p, we estimate the change in volume for the dimerization of actin, Delta V-dim, to be -307 +/- 10 ml/mol at 279 K. The change of T-p with pressure dT(p)/dP=(0.3015 +/- 0.0009) K/MPa=(30.15 +/- 0.09) mK/atm. The phi(T,P) data at higher T indicate the change in volume on propagation, Delta V-prop, to be +401 +/- 48 ml/mol at 301 K. The phi(t) measurements yield initial relaxation times r(p)(T) that reflect the behavior of phi(T) and support the presence of a depolymerization temperature. We also measured the density of polymerizing actin with a vibrating tube density meter, the results of which confirm that the data from this instrument are affected by viscosity changes and can be erroneous. (c) 2005 American Institute of Physics.