MOUSE POLYCLONAL ANTIBODIES TO UDP-N-ACETYLGLUCOSAMINE - LYSOSOMAL-ENZYME N-ACETYLGLUCOSAMINE-1-PHOSPHOTRANSFERASE FROM ACANTHAMOEBA-CASTELLANII

Mice were immunized with preparations of UDP-N-acetylglucosamine:lysos omal enzyme ne-1-phosphotransferase(GlcNAc-phosphotransferase) purifie d more than 10,000 fold. Specific antibodies for Acanthamoeba castella nii proteins and with high affinity for this enzyme were obtained. Cru de and highly purified GlcNAc-phosphotransferase preparations were chr omatographed on an exclusion molecular column, in which the enzyme pre sented elution volumes that depended on the preparations. The proteins of the eluted fractions were separated by electrophoresis and immunob lots developed with the mouse antiserum. Analyses of the proteins that reacted with the antibodies and whose intensity profiles correlated w ith GlcNAc-phosphotransferase activity, showed that two proteins of 71 kDa and 82 kDa are probably subunits or subunit fragments of the GlcN Ac-phosphotransferase.