Evidence for extensive subcellular organization of asparagine-linked oligosaccharide processing and lysosomal enzyme phosphorylation.

Membranes prepared from mouse lymphoma BW5147.3 cells and P388D1 macrophages were fraction-ated on a continuous sucrose gradient and assayed for enzymes involved in the processing of asparagine-linked oligosaccharides. The order in which these en-zymes distributed from dense to light membranes cor-related with the established sequence of events in gly-coprotein biosynthesis. A number of enzymes which have been previously localized to the Golgi separated into four regions on the gradient. UDP-N-acetylgluco-samine:lysosomal enzyme N-acetylglucosamine-l-phosphotransferase, the enzyme which catalyzes the selective phosphorylation of the high mannose oligo-saccharides of lysosomal enzymes, was present in the densest membranes. N-Acetylglucosamine l-phospho-