Nerve growth factor-inducible large external (NILE) glycoprotein: studies of a central and peripheral neuronal marker.
The PC12 clone of pheochromocytoma cells undergoes neuronal differentiation in the presence of nerve growth factor (NGF). Concomitant with this is a significant induction in the incorporation of radiolabeled fucose or glucosamine into a 230,000-dalton cell surface glycoprotein named the NGF-Inducible Large External, or NILE, glycoprotein (GP) (McGuire, J. C., L. A. Greene, and A. V. Furano (1978) Cell 15: 357-365). In the current studies NILE GP was purified from PC12 cells using wheat germ agglutinin-agarose affinity chromatography and SDS-polyacrylamide gel electrophoresis (PAGE). Polyclonal antisera were raised against purified NILE GP and were found to selectively immunoprecipitate a single 230,000-dalton protein from detergent extracts of PC12 cells metabolically labeled with either [3H]fucose, [3H]glucosamine, or [35S]methionine. These antisera stained the surfaces of PC12 cells by indirect immunofluorescence and were cytotoxic to PC12 cells in the presence of complement. Limited treatment of PC12 cells with either trypsin or pronase produced a fucosylated 90,000-dalton immunoreactive fragment of NILE GP which remained in the membrane. Using quantitative immunoelectrophoresis, the action of NGF on NILE GP was represent an increase in the amount of protein, rather than a selective increase in carbohydrate incorporation. Immunofluorescent staining of primary cell cultures and tissue whole mounts revealed that immunologically cross-reactive NILE GP appears to be expressed on the cell surfaces (somas and neurites) of most if not all peripheral and central neurons examined. Immunoprecipitation of radiolabeled cultures showed that the cross-reactive material had an apparent molecular weight by SDS-PAGE of 225,000 to 230,000 in the peripheral nervous system and 200,000 to 210,000 in the central nervous system. NILE-cross-reactive material was also found to a small extent on Schwann cell surfaces, but not at all on a variety of other cell types. These results suggest that immunoreactive NILE GP is distributed widely and selectively on neural cell surfaces.
D000310:Adrenal Gland Neoplasms / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002490:Central Nervous System / Q000378:metabolism / Q000502:physiology; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000302:isolation & purification*; D007106:Immune Sera / Q000276:immunology; D007122:Immunoelectrophoresis; D008970:Molecular Weight; D009414:Nerve Growth Factors / Q000494:pharmacology / Q000502:physiology*; D039842:Neural Cell Adhesion Molecule L1; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D010525:Peripheral Nerves / Q000378:metabolism / Q000502:physiology; D010673:Pheochromocytoma / Q000378:metabolism; D051381:Rats
D006023:Glycoproteins; D007106:Immune Sera; D009414:Nerve Growth Factors; D039842:Neural Cell Adhesion Molecule L1