The neurotrophin receptor, gp75, forms a complex with the receptor tyrosine kinase TrkA
Authors
Ross, Alonzo H.Daou, Marie-Claire
McKinnon, Christine A.
Condon, Peter J.
Lachyankar, Mahesh B.
Stephens, Robert M.
Kaplan, David R.
Wolf, David E.
UMass Chan Affiliations
Department of Biochemistry and Molecular PharmacologyGraduate School of Biomedical Sciences
Document Type
Journal ArticlePublication Date
1996-03-01Keywords
Animals; Cell Compartmentation; Cells, Cultured; Fluorescent Antibody Technique; Protein Binding; Proto-Oncogene Proteins; Receptor Protein-Tyrosine Kinases; Receptor, Nerve Growth Factor; Receptor, trkA; Receptors, Nerve Growth Factor; Recombinant Proteins; SpodopteraLife Sciences
Medicine and Health Sciences
Metadata
Show full item recordAbstract
The high-affinity NGF receptor is thought to be a complex of two receptors , gp75 and the tyrosine kinase TrkA, but direct biochemical evidence for such an association had been lacking. In this report, we demonstrate the existence of such a gp75-TrkA complex by a copatching technique. Gp75 on the surface of intact cells is patched with an anti-gp75 antibody and fluorescent secondary antibody, the cells are then fixed to prevent further antibody-induced redistributions, and the distribution of TrkA is probed with and anti-TrkA antibody and fluorescent secondary antibody. We utilize a baculovirus-insect cell expression of wild-type and mutated NGF receptors. TrkA and gp75 copatch in both the absence and presence of NGF. The association is specific, since gp75 does not copatch with other tyrosine kinase receptors, including TrkB, platelet-derived growth factor receptor-beta, and Torso (Tor). To determine which domains of TrkA are required for copatching, we used a series of TrkA-Tor chimeric receptors and show that the extracellular domain of TrkA is sufficient for copatching with gp75. A chimeric receptor with TrkA transmembrane and intracellular domains show partial copatching with gp75. Deletion of the intracellular domain of gp75 decreases but does not eliminate copatching. A point mutation which inactivates the TrkA kinase has no effect on copatching, indicating that this enzymatic activity is not required for association with gp75. Hence, although interactions between the gp75 and TrkA extracellular domains are sufficient for complex formation, interactions involving other receptor domains also play a role.Source
J Cell Biol. 1996 Mar;132(5):945-53.
DOI
10.1083/jcb.132.5.945Permanent Link to this Item
http://hdl.handle.net/20.500.14038/32454PubMed ID
8603925Related Resources
Distribution License
http://creativecommons.org/licenses/by-nc-sa/4.0/ae974a485f413a2113503eed53cd6c53
10.1083/jcb.132.5.945
Scopus Count
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Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by-nc-sa/4.0/
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