Biosynthesis and membrane topography of the neural cell adhesion molecule L1.
The biosynthesis and membrane topography of the neural cell adhesion molecule L1 have been studied in cerebellar cell cultures by metabolic labeling and immunoprecipitation. Pulse and pulse-chase experiments with [35S]methionine show that L1 is synthesized in its high mol. wt. form, the 200 kd component. The lower mol. wt. components with 40, 80 and 140 K apparent mol. wts. can be generated by proteolysis in intact cellular membranes. Peptide maps generated by protease treatment of L1 isolated from adult mouse brain show that the 80 and 140 kd components are related to the 200 kd component, but not to each other. The 200, 80 and 40 kd components can be biosynthetically phosphorylated. The 140 kd component is not phosphorylated and not released from the surface membrane during tryspinization. The phosphorylated amino acid is serine. In the presence of tunicamycin the 200 kd component is synthesized as a 150 kd protein. Pulse-chase experiments in the presence of tunicamycin indicate that the carbohydrate moieties are predominantly N-glycosidically linked and that the contribution of O-glycosylation is minimal. The carbohydrate moieties are of the complex type as shown by treatment with endoglycosidase H. Since monensin inhibits processing of the carbohydrate moieties, the 200 kd component appears to be transported to the surface membrane via the Golgi apparatus.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000831:Animals, Newborn; D000954:Antigens, Surface / Q000032:analysis* / Q000235:genetics; D015815:Cell Adhesion Molecules; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D002531:Cerebellum / Q000196:embryology / Q000378:metabolism*; D005260:Female; D005643:Fucose / Q000378:metabolism; D005944:Glucosamine / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D010446:Peptide Fragments / Q000032:analysis; D010710:Phosphates / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D011247:Pregnancy; D013462:Sulfur Radioisotopes; D014316:Tritium; D014357:Trypsin
D000596:Amino Acids; D000954:Antigens, Surface; D015815:Cell Adhesion Molecules; D010446:Peptide Fragments; D010710:Phosphates; D010761:Phosphorus Radioisotopes; D013462:Sulfur Radioisotopes; D014316:Tritium; D005643:Fucose; D008715:Methionine; D014357:Trypsin; D005944:Glucosamine