Mechanism of ATP hydrolysis by polymeric actin.
The lag between polymerization of actin and ATP hydrolysis in actin filaments was analyzed in terms of the mechanism of the hydrolysis reaction. Under the experimental conditions (100 mM KCl and 1 mM MgCl2, or without KCl, 1 mM MgCl2 and 0.4 mM EGTA, 25 degrees C) ATP hydrolysis lagged behind polymerization by about 100 s independently of the concentration of polymerizing filament ends and of the actin monomer concentration. Three models of ATP hydrolysis were compared to experimental data: (i) Random ATP hydrolysis, ATP is assumed to be hydrolyzed at a rate that is independent of the type of the nucleotide bound to adjacent filament subunits. (ii) Cooperative hydrolysis, the rate of ATP hydrolysis is thought to depend on the type of nucleotide bound to adjacent subunits. (iii) Sequential hydrolysis, ATP is assumed to be hydrolyzed only at the interface between ATP-subunits and ADP-subunits. The model of sequential ATP hydrolysis could be excluded. The results were in agreement with random or cooperative ATP hydrolysis. The differences of the rates of ATP hydrolysis by a random or cooperative mechanism are so small that based on the experimental results no distinction between these two mechanisms could be made. All available evidence points towards a mechanism of ATP hydrolysis in which several or perhaps many interfaces between ATP- and ADP-subunits are formed within a filament.
Ohm
T
T
Institute of Physiological Chemistry, Ruhr-University, Bochum, Germany.
Wegner
A
A
D000199:Actins / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004533:Egtazic Acid / Q000494:pharmacology; D006868:Hydrolysis; D007700:Kinetics; D015636:Magnesium Chloride / Q000494:pharmacology; D011108:Polymers / Q000378:metabolism*; D011189:Potassium Chloride / Q000494:pharmacology; D011817:Rabbits
D000199:Actins; D011108:Polymers; D015636:Magnesium Chloride; D004533:Egtazic Acid; D000244:Adenosine Diphosphate; D011189:Potassium Chloride; D000255:Adenosine Triphosphate