Impact of profilin on actin-bound nucleotide exchange and actin polymerization dynamics.
We have investigated the effects of profilin on nucleotide binding to actin and on steady state actin polymerization. The rate constants for the dissociation of ATP and ADP from monomeric Mg-actin at physiological conditions are 0.003 and 0.009 s-1, respectively. Profilin increases these dissociation rate constants to 0.08 s-1 for MgATP-actin and 1.4 s-1 for MgADP-actin. Thus, profilin can increase the rate of exchange of actin-bound ADP for ATP by 140-fold. The affinity of profilin for monomeric actin is found to be similar for MgATP-actin and MgADP-actin. Continuous sonication was used to allow study of solutions having sustained high filament end concentrations. During sonication at steady state, F-actin depolymerizes toward the critical concentration of ADP-actin [Pantaloni, D., et al. (1984)J. Biol. Chem. 259, 6274-6283], our analysis indicates that under these conditions a significant number of filaments contain terminal ADP-actin subunits. Addition of profilin to this system increases the polymer concentration and increases the steady state ATPase activity during sonication. These data are explained by the fast exchange of ATP for ADP on the profilin-ADP-actin complex, resulting in rapid ATP-actin regeneration. An important function of profilin may be to provide the growing ends of filaments with ATP-actin during periods when the monomer cycling rate exceeds the intrinsic nucleotide exchange rate of monomeric actin.
Selden
L A
LA
Research Service, Stratton VA Medical Center, Albany, New York 12208, USA.
Kinosian
H J
HJ
Estes
J E
JE
Gershman
L C
LC
D000199:Actins / Q000737:chemistry* / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D003285:Contractile Proteins; D006868:Hydrolysis; D007700:Kinetics; D008432:Mathematical Computing; D008840:Microfilament Proteins / Q000502:physiology*; D009994:Osmolar Concentration; D011108:Polymers / Q000737:chemistry; D051304:Profilins; D011485:Protein Binding; D012996:Solutions; D013010:Sonication; D013816:Thermodynamics
D000199:Actins; D003285:Contractile Proteins; D008840:Microfilament Proteins; D011108:Polymers; D051304:Profilins; D012996:Solutions; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate