Mechanism of S100 protein-dependent inhibition of glial fibrillary acidic protein (GFAP) polymerization

Abstract

S100 protein, a subfamily of Ca(2+)-binding proteins of the EF-hand type, was recently shown to bind to and to inhibit the polymerization of the glial fibrillary acidic protein (GFAP), the intermediate filament component of astroglial cells, in the presence of micromolar levels of Ca2+ (J. Biol. Chem. 268, 12669-12674). By a sedimentation assay and viscometry we show here that S100 protein interferes with the very early steps of GFAP polymerization (nucleation) and with the GFAP polymer growth, thereby retarding the onset of GFAP assembly, reducing the rate and the extent of GFAP assembly, and increasing the critical concentration of GFAP assembly. Moreover, S100 protein disassembles preformed glial filaments. All the above effects can be explained by sequestration of soluble GFAP by S100 protein, as also indicated by the stoichiometry of S100 protein binding to GFAP and of S100 protein effects on GFAP assembly. Our data suggest that S100 protein might serve the function of avoiding excess GFAP polymerization and might participate in remodeling of glial filaments following elevation of the intracellular free Ca2+ concentration. Also, our data lend support to the notion that intermediate filaments are dynamic cytoskeleton structures that assemble and disassemble, and to the existence of cytoplasmic factors implicated in the regulation of the state of assembly of intermediate filaments.