Jager_1990_Biochim.Biophys.Acta_1039_367

Dimeric acetylcholinesterase is anchored in the cell membrane by a glycosyl-phosphatidylinositol attached to the C-terminus of the protein. The complex glycan contains an antigenic epitope, the cross-reacting determinant (CRD), which is only revealed after removal of the diradylglycerol by phosphatidylinositol-specific phospholipase C (PI-PLC) but is cryptic in the amphiphilic form. Polyclonal antibodies were raised against the CRD of vertebrate acetylcholinesterase. The purified anti-CRD antibodies recognized only the PI-PLC treated hydrophilic forms of acetylcholinesterase from bovine erythrocytes and Torpedo, and of variant surface glycoprotein from trypanosomes but not the corresponding amphiphilic proteins. Competition experiments showed that inositol-1,2-cyclic phosphate and glucosamine inhibited the binding of the antibodies to the CRD. Furthermore, binding of the anti-CRD antibodies to acetylcholinesterase containing N-methylated glucosamine was markedly reduced. The amphiphilic N-methylated enzyme is less sensitive to digestion with PI-PLC than the non-methylated form. From our results we conclude that inositol-1,2-cyclic phosphate and glucosamine, especially the free amine group of this residue, contribute significantly to the epitope recognized by the anti-CRD antibodies.