H2O2-treated actin: assembly and polymer interactions with cross-linking proteins.
During inflammation, hydrogen peroxide, produced by polymorphonuclear leukocytes, provokes cell death mainly by disarranging filamentous (polymerized) actin (F-actin). To show the molecular mechanism(s) by which hydrogen peroxide could alter actin dynamics, we analyzed the ability of H2O2-treated actin samples to polymerize as well as the suitability of actin polymers (from oxidized monomers) to interact with cross-linking proteins. H2O2-treated monomeric (globular) actin (G-actin) shows an altered time course of polymerization. The increase in the lag phase and the lowering in both the polymerization rate and the polymerization extent have been evidenced. Furthermore, steady-state actin polymers, from oxidized monomers, are more fragmented than control polymers. This seems to be ascribable to the enhanced fragility of oxidized filaments rather than to the increase in the nucleation activity, which markedly falls. These facts; along with the unsuitability of actin polymers from oxidized monomers to interact with both filamin and alpha-actinin, suggest that hydrogen peroxide influences actin dynamics mainly by changing the F-actin structure. H2O2, via the oxidation of actin thiols (in particular, the sulfhydryl group of Cys-374), likely alters the actin C-terminus, influencing both subunit/subunit interactions and the spatial structure of the binding sites for cross-linking proteins in F-actin. We suggest that most of the effects of hydrogen peroxide on actin could be explained in the light of the "structural connectivity," demonstrated previously in actin.
DalleDonne
I
I
Department of Biology, University of Milan, Italy.
Milzani
A
A
Colombo
R
R
D000185:Actinin / Q000378:metabolism; D000199:Actins / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D002645:Chickens; D003285:Contractile Proteins / Q000378:metabolism; D003432:Cross-Linking Reagents; D064448:Filamins; D005895:Gizzard, Avian; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007700:Kinetics; D008027:Light; D046911:Macromolecular Substances; D008840:Microfilament Proteins / Q000378:metabolism; D008854:Microscopy, Electron; D018482:Muscle, Skeletal / Q000378:metabolism; D009130:Muscle, Smooth / Q000378:metabolism; D010084:Oxidation-Reduction; D011817:Rabbits; D012542:Scattering, Radiation; D014783:Viscosity
D000199:Actins; D003285:Contractile Proteins; D003432:Cross-Linking Reagents; D064448:Filamins; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D000185:Actinin; D006861:Hydrogen Peroxide