title: Studies of the biosynthesis of the mannose 6-phosphate receptor in receptor-positive and -deficient cell lines.
abstract: Phosphomannosyl residues present on lysosomal enzymes are specifically recognized by the mannose 6-phosphate receptor protein. This interaction results in the selective targeting of lysosomal enzymes to lysosomes. While this pathway is operative in many cell types, we have found four cultured cell lines that are deficient in the ability to bind lysosomal enzymes containing phosphomannosyl residues to their intracellular or surface membranes (Gabel, C., D. Goldberg, and S. Kornfeld, 1983, Proc. Natl. Acad. Sci. USA, 80:775-779). These cells appear to segregate lysosomal enzymes by an alternate intracellular pathway. To determine the basis for the lack of mannose 6-phosphate receptor activity in these cell lines, we studied the biosynthesis of the receptor in receptor-positive (BW5147) and receptor-deficient (P388D1 and MOPC 315) cells. The cells were labeled with [2-3H]mannose or [35S]methionine and the receptor was immunoprecipitated with an antireceptor antiserum. BW5147 cells synthesize a receptor protein whose size increases after translation/glycosylation. MOPC 315 cells produce an apparently normal receptor and degrade it rapidly. P388D1 cells fail to synthesize any detectable receptor. The receptor from BW5147 and MOPC 315 cells is a glycoprotein with both high mannose and complex asparagine-linked oligosaccharides. The complex-type units become fully sialylated and remain so during long periods of chase.
mesh_terms: D000818:Animals; D002460:Cell Line; D006020:Glycopeptides / Q000302:isolation & purification; D006600:Hexosephosphates / Q000378:metabolism*; D007941:Leukemia P388 / Q000378:metabolism; D008223:Lymphoma / Q000378:metabolism; D008360:Mannosephosphates / Q000378:metabolism*; D051379:Mice; D010954:Plasmacytoma / Q000378:metabolism; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D006020:Glycopeptides; D006600:Hexosephosphates; D008360:Mannosephosphates; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface; C027693:mannose-6-phosphate