An intermediate state of G-actin between native and denatured: polymerization rate decreases but extent of polymerization remains unchanged.
The rate of actin polymerization gradually decreased without changing the final level of polymerization, when incubated in the presence of 0.2 mM ATP at pH 8.0 and 25 degrees C. This change was much faster in Mg2+-actin than Ca2+-actin, and Mg2+-actin became denatured and unpolymerizable on prolonged incubation. The drop in the polymerization rate was due both to weakened nucleation and a slowed elongation rate in the incubated actin. The change in the polymerization rate was partially reversible by storing the sample at 0 degrees C. When the rate of polymerization dropped markedly on prolonged incubation, a gel filtration profile showed that Ca2+-actin existed as monomer not as oligomer. On the other hand, Mg2+-actin formed dimers, and other oligomers, as revealed by crosslinking analysis. There were changes in fluorescence intensities due to tyrosine and/or tryptophan residues of the actin molecule, and in difference absorption spectra, suggesting that conformational changes intermediate between native and denatured states occurred during incubation.
D000199:Actins / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D009132:Muscles / Q000378:metabolism; D011489:Protein Denaturation; D011817:Rabbits; D013997:Time Factors
D000199:Actins; D046911:Macromolecular Substances; D000255:Adenosine Triphosphate; D008274:Magnesium; D002118:Calcium