Association of nerve growth factor receptors with the triton X-100 cytoskeleton of PC12 cells.
Triton X-100 solubilizes membranes of PC12 cells and leaves behind a nucleus and an array of cytoskeletal filaments. Nerve growth factor (NGF) receptors (10% of those found in intact cells) are associated with this Triton X-100-insoluble residue. Two classes of NGF receptors are found on PC12 cells which display rapid and slow dissociating kinetics. Although rapidly dissociating binding is predominant (greater than 75%) in intact cells, the majority of binding to the Triton X-100 cytoskeleton is slowly dissociating (greater than 75%). Rapidly dissociating NGF binding on intact cells can be converted to a slowly dissociating form by the plant lectin wheat germ agglutinin (WGA). This lectin also increases the number of receptors which associate with the Triton X-100 cytoskeleton by more than 10-fold. 125I-NGF bound to receptors can be visualized by light microscopy autoradiography in Triton X-100-insoluble residues of cell bodies, as well as growth cones and neurites. The WGA-induced association with the cytoskeleton, however, is not specific for the NGF receptor, since greater than 90% of cell surface glycoprotein receptors for WGA become associated with Triton X-100-insoluble material at lectin concentrations greater than 33 micrograms/ml. Concentrations of WGA which change the Triton X-100 solubility of membrane glycoproteins are similar to those required to alter the kinetic state of the NGF receptor. Both events may be related to the crossbridging of cell surface proteins induced by this multivalent lectin.
D000310:Adrenal Gland Neoplasms / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D003584:Cytological Techniques; D003599:Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D007457:Iodine Radioisotopes; D007700:Kinetics; D008565:Membrane Proteins / Q000032:analysis; D008855:Microscopy, Electron, Scanning; D009414:Nerve Growth Factors / Q000378:metabolism; D017830:Octoxynol; D010673:Pheochromocytoma / Q000378:metabolism* / Q000473:pathology; D011092:Polyethylene Glycols; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017475:Receptors, Nerve Growth Factor; D013997:Time Factors; D014018:Tissue Distribution
D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D009414:Nerve Growth Factors; D011956:Receptors, Cell Surface; D017475:Receptors, Nerve Growth Factor; D011092:Polyethylene Glycols; D017830:Octoxynol