Nucleotide exchange and rheometric studies with F-actin prepared from ATP- or ADP-monomeric actin.
It has recently been reported that polymer actin made from monomer containing ATP (ATP-actin) differed in EM appearance and rheological characteristics from polymer made from ADP-containing monomers (ADP-actin). Further, it was postulated that the ATP-actin polymer was more rigid due to storage of the energy released by ATP hydrolysis during polymerization (Janmey et al. 1990. Nature 347:95-99). Electron micrographs of our preparations of ADP-actin and ATP-actin polymers show no major differences in appearance of the filaments. Moreover, the dynamic viscosity parameters G' and G" measured for ATP-actin and ADP-actin polymers are very different from those reported by Janmey et al., in absolute value, in relative differences, and in frequency dependence. We suggest that the relatively small differences observed between ATP-actin and ADP-actin polymer rheological parameters could be due to small differences either in flexibility or, more probably, in filament lengths. We have measured nucleotide exchange on ATP-actin and ADP-actin polymers by incorporation of alpha-32P-ATP and found it to be very slow, in agreement with earlier literature reports, and in contradiction to the faster exchange rates reported by Janmey et al. This exchange rate is much too slow to cause "reversal" of ADP-actin polymer ATP-actin polymer as reported by Janmey et al. Thus our results do not support the notion that the energy of actin-bound ATP hydrolysis is trapped in and significantly modifies the actin polymer structure.
Newman
J
J
Department of Physics, Union College, Schenectady, New York 12308.
Zaner
K S
KS
Schick
K L
KL
Gershman
L C
LC
Selden
L A
LA
Kinosian
H J
HJ
Travis
J L
JL
Estes
J E
JE
D000199:Actins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D000244:Adenosine Diphosphate / Q000031:analogs & derivatives* / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000737:chemistry; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D004548:Elasticity; D008854:Microscopy, Electron; D011817:Rabbits; D012212:Rheology; D013816:Thermodynamics; D014783:Viscosity
C042634:ADP-G-actin; C022920:ATP-G-actin; D000199:Actins; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate