Effects of solution crowding on actin polymerization reveal the energetic basis for nucleotide-dependent filament stability

JOURNAL OF MOLECULAR BIOLOGY

卷:378

Article

Frederick, Kendra B.¹  Sept, David²  De La Cruz, Enrique M.¹ 

[1] Yale Univ, Dept Mol Biophys & Biochem, New Haven, CT 06520 USA

[2] Washington Univ, Dept Biomed Engn, St Louis, MO 63130 USA

关键词: actin;    crowding theory;    nucleotide;    thermodynamics;    kinetics;   

DOI  :  10.1016/j.jmb.2008.02.022

来源: Elsevier

【 摘 要 】

Actin polymerization is a fundamental cellular process involved in cell structure maintenance, force generation, and motility. Phosphate release from filament subunits following ATP hydrolysis destabilizes the filament lattice and increases the critical concentration (C-c) for assembly. The structural differences between ATP- and ADP-actin are still debated, as well as the energetic factors that underlie nucleotide-dependent filament stability, particularly under crowded intracellular conditions. Here, we investigate the effect of crowding agents on ATP- and ADP-actin polymerization and find that ATP-actin polymerization is largely unaffected by solution crowding, while crowding agents lower the C-c of ADP-actin in a concentration-dependent manner. The stabilities of ATP- and ADP-actin filaments are comparable in the presence of physiological amounts (similar to 30% w/v) and types (sorbitol) of low molecular weight crowding agents. Crowding agents act to stabilize ADP-F-actin by slowing subunit dissociation. These observations suggest that nucleotide hydrolysis and phosphate release per se do not introduce intrinsic differences in the in vivo filament stability. Rather, the preferential disassembly of ADP-actin filaments in cells is driven through interactions with regulatory proteins. Interpretation of the experimental data according to osmotic stress theory implicates water as an allosteric regulator of actin activity and hydration as the molecular basis for nucleotide-dependent filament stability. (C) 2008 Elsevier Ltd. All rights reserved.

【 授权许可】

Free