Thermodynamics of actin polymerization; influence of the tightly bound divalent cation and nucleotide.
Previous work by this laboratory has shown that the tightly bound divalent cation of actin affects the enthalpy of the polymerization reaction for ATP-actin (Selden et al. (1986) J. Muscle Res. Cell Motil. 7, 215-224). In the present study, we have measured the temperature dependence of polymerization for actin containing ATP or ADP as the bound nucleotide and Mg2+ or Ca2+ (Mg-actin or Ca-actin) as the tightly bound divalent cation. In contrast to the marked effect of the tightly bound divalent cation on enthalpy and entropy changes for the polymerization of ATP-actin, ADP-actin polymerization is affected very little by the tightly bound divalent cation. The Arrhenius and van't Hoff plots for polymerization of Ca-ATP-, Mg-ADP- and Ca-ADP-actin were found to be non-linear. The free energy data for actin polymerization have been analyzed as a second order function of absolute temperature (Osborne et al. (1976) Biochemistry 15, 317-320). The values of the enthalpy change and activation enthalpy change for Ca-ATP-, Mg-ADP- and Ca-ADP-actin polymerization were found to be temperature-dependent, in contrast to those for Mg-ATP-actin, which were nearly constant over the temperature range studied. These results suggest that (1) polymerization of actin which does not contain both Mg2+ and ATP may be a multi-step reaction including a rate-limiting step and (2) Mg-ATP-actin has a unique conformation which enhances its ability to polymerize.
Kinosian
H J
HJ
Department of Physiology and Cell Biology, Albany Medical College, NY.
Selden
L A
LA
Estes
J E
JE
Gershman
L C
LC
D000199:Actins / Q000737:chemistry*; D000244:Adenosine Diphosphate / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D002118:Calcium / Q000737:chemistry; D002413:Cations, Divalent / Q000737:chemistry*; D005454:Fluorescence Polarization; D008274:Magnesium / Q000737:chemistry; D009711:Nucleotides / Q000737:chemistry*; D011108:Polymers; D013696:Temperature; D013816:Thermodynamics
D000199:Actins; D002413:Cations, Divalent; D009711:Nucleotides; D011108:Polymers; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D008274:Magnesium; D002118:Calcium