Abstract

The major goal of the proposed research is to achieve a detailed biochemical characterization of the nerve growth factor receptor, in order to help elucidate the molecular mechanism of NGF action. The NGF receptor will be purified from human melanoma cells and the biochemical properties of the NGF receptor will be investigated. In particular, biochemical heterogeneity of the receptor which might account for observed heterogeneity in NGF binding properties will be sought, and the possibility that the receptor has a protein kinase activity, and that it may physically interact with the cytoskeleton will be explored. Monoclonal antibodies to the NGF receptor will be produced and used as an aid for receptor purification. DNA-mediated gene transfer will be used to obtain integration and stable expression of the human NGF receptor (ngfr) gene in mouse cells. After serial passage of the human ngfr gene through mouse cells by transfection, the human ngfr gene will be molecularly cloned by isolation of lambda phage recombinants containing human DNA. Nucleotide sequence data will be obtained for the cloned gene and used in conjunction with biochemical data to achieve an understanding of NGF receptor structure, and its possible relationship to the structure and function of other polypeptide hormone receptors.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS023343-02
Application #
3406683
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1985-07-01
Project End
1987-06-30
Budget Start
1986-07-01
Budget End
1987-06-30
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Washington
Department
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195

  Publications

Garden, G A; Hartlage-Rubsamen, M; Rubel, E W et al. (1995) Protein masking of a ribosomal RNA epitope is an early event in afferent deprivation-induced neuronal death. Mol Cell Neurosci 6:293-310
Borson, S; Schatteman, G; Claude, P et al. (1994) Neurotrophins in the developing and adult primate adenohypophysis: a new pituitary hormone system? Neuroendocrinology 59:466-76
Garden, G A; Canady, K S; Lurie, D I et al. (1994) A biphasic change in ribosomal conformation during transneuronal degeneration is altered by inhibition of mitochondrial, but not cytoplasmic protein synthesis. J Neurosci 14:1994-2008
Henry, M A; Westrum, L E; Bothwell, M et al. (1994) Electron microscopic localization of nerve growth factor receptor (p75)-immunoreactivity in pars caudalis/medullary dorsal horn of the cat. Brain Res 642:137-45
Patterson, S L; Grady, M S; Bothwell, M (1993) Nerve growth factor and a fibroblast growth factor-like neurotrophic activity in cerebrospinal fluid of brain injured human patients. Brain Res 605:43-9
Schatteman, G C; Langer, T; Lanahan, A A et al. (1993) Distribution of the 75-kD low-affinity nerve growth factor receptor in the primate peripheral nervous system. Somatosens Mot Res 10:415-32
von Bartheld, C S; Bothwell, M (1993) Development of the mesencephalic nucleus of the trigeminal nerve in chick embryos: target innervation, neurotrophin receptors, and cell death. J Comp Neurol 328:185-202
Taiji, M; Taiji, K; Deyerle, K L et al. (1992) Basic fibroblast growth factor enhances nerve growth factor receptor gene promoter activity in human neuroblastoma cell line CHP100. Mol Cell Biol 12:2193-202
Schecterson, L C; Bothwell, M (1992) Novel roles for neurotrophins are suggested by BDNF and NT-3 mRNA expression in developing neurons. Neuron 9:449-63
von Bartheld, C S; Bothwell, M (1992) Development and distribution of noradrenergic and cholinergic neurons and their trophic phenotypes in the avian ceruleus complex and midbrain tegmentum. J Comp Neurol 320:479-500

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