Modifications of neurofilament proteins by possible metabolites of allyl chloride in vitro

Chronic exposure to allyl chloride (ALL) is known to produce a central-peripheral distal axonopathy. In relation to the mechanism(s), the present study was conducted to examine the abilities of ALL and its putative metabolites, i.e., epichlorohydrin, glycerol alpha-monochlorohydrin, allyl alcohol and acrolein to crosslink proteins in vitro. Neurofilament-riched cytoskeletal proteins (1 mg/ml) and ovalbumin (10 mg/ml) were incubated with 160 mM tested chemicals except for acrolein at 0.5 mM and 1 mM. Time-dependent studies by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that not only ALL, but also acrolein and epichlorohydrin exerted chemical modifications on axonal cytoskeletal proteins; while only acrolein-treated ovalbumin could manifest evidence of polymerization of the protein. Immunoblotting of PAGE-separated proteins confirmed that the high molecular weight proteins on the top of SDS-PAGEs were NF antigen-contained covalent cross-linked materials.