Arevalo, Juan C
(Instituto de Microbiologia Bioquimica, Departamento de Microbiologia y Genetica, CSIC Universidad de Salamanca, 37007 Salamanca, Spain)
,
Conde, Blanca
(Departamento de Ciencias Morfologicas, Universidad de Zaragoza, Spain)
,
Hempstead, Barbara I
(Hematology)
,
Chao, Moses V
(Molecular Neurobiology Program, Skirball Institute, Departments of Cell Biology and Physiology and Neuroscience, New York University Medical Center, New York, NY 10016, USA)
,
Martín-Zanca, Dionisio
(Instituto de Microbiologia Bioquimica, Departamento de Microbiologia y Genetica, CSIC Universidad de Salamanca, 37007 Salamanca, Spain)
,
Pérez, Pilar
(Instituto de Microbiologia Bioquimica, Departamento de Microbiologia y Genetica, CSIC Universidad de Salamanca, 37007 Salamanca, Spain)
The TrkA NGF receptor extracellular region contains three leucine repeats flanked by cysteine clusters and two immunoglobulin-like domains that are required for specific ligand binding. Deletion of the immunoglobulin-like domains abolishes NGF binding and causes ligand independent activation of the ...
The TrkA NGF receptor extracellular region contains three leucine repeats flanked by cysteine clusters and two immunoglobulin-like domains that are required for specific ligand binding. Deletion of the immunoglobulin-like domains abolishes NGF binding and causes ligand independent activation of the receptor. Here we report a specific mutation that increases the binding affinity of the TrkA receptor for NGF. A change of proline 203 to alanine (P203A) in the linker region between the leucine repeats and the first Ig-like domain increased NGF binding by decreasing the ligand rate of dissociation. This mutated receptor was appropriately expressed on the cell surface and promoted ligand-independent neurite outgrowth in PC12nnr5 cells. The mutant receptor was capable of spontaneous dimerization and was constitutively phosphorylated in the absence of ligand. Moreover, expression of TrkA-P203A receptor in fibroblasts induced DNA synthesis and transformation and generated tumours in nude mice. These data suggest that domains outside of the immunoglobulin-like structure contribute to ligand binding and constitutive activation of Trk receptors.
The TrkA NGF receptor extracellular region contains three leucine repeats flanked by cysteine clusters and two immunoglobulin-like domains that are required for specific ligand binding. Deletion of the immunoglobulin-like domains abolishes NGF binding and causes ligand independent activation of the receptor. Here we report a specific mutation that increases the binding affinity of the TrkA receptor for NGF. A change of proline 203 to alanine (P203A) in the linker region between the leucine repeats and the first Ig-like domain increased NGF binding by decreasing the ligand rate of dissociation. This mutated receptor was appropriately expressed on the cell surface and promoted ligand-independent neurite outgrowth in PC12nnr5 cells. The mutant receptor was capable of spontaneous dimerization and was constitutively phosphorylated in the absence of ligand. Moreover, expression of TrkA-P203A receptor in fibroblasts induced DNA synthesis and transformation and generated tumours in nude mice. These data suggest that domains outside of the immunoglobulin-like structure contribute to ligand binding and constitutive activation of Trk receptors.
참고문헌 (35)
10.1128/MCB.20.16.5908-5916.2000 Arevalo JC, Conde B, Hempstead BL, Chao MV, Martin-Zanca D and Perez P. . 2000 Mol. Cell. Biol. 20: 5908-5916.
Ezoe K, Holmes SA, Ho L, Bennett CP, Bolognia JL, Brueton L, Burn J, Falabella R, Gatto EM, Issii N, et al. . 1995 Am. J. Hum. Gen. 56: 58-66.
10.1002/(SICI)1098-2264(199706)19:2<112::AID-GCC7>3.0.CO;2-1 Greco A, Miranda C, Pagliardini S, Fusetti L, Bongarzone I and Pierotti MA. . 1997 Genes Chromosomes Cancer 19: 112-123.
10.1038/ng1194-269 Muenke M, Schell U, Hehr A, Robin NH, Losken HW, Schinzel A, Pulleyn LJ, Rutland P, Reardon W, Malcolm S and Winter RM. . 1994 Nature Genet. 8: 269-274.
10.1038/ng0295-165 Wilkie AOM, Slaney SF, Oldrigde M, Poole MD, Ashworth GJ, Hockley AD, Hayward RD, David DJ, Pulleyn L and Rutland P. . 1995 Nature Genet. 9: 165-172.
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