Binding characteristics of brain-derived neurotrophic factor to its receptors on neurons from the chick embryo.
Brain-derived neurotrophic factor (BDNF), a protein known to support the survival of embryonic sensory neurons and retinal ganglion cells, was derivatized with 125I-Bolton-Hunter reagent and obtained in a biologically active, radioactive form (125I-BDNF). Using dorsal root ganglion neurons from chick embryos at 9 d of development, the basic physicochemical parameters of the binding of 125I-BDNF with its receptors were established. Two different classes of receptors were found, with dissociation constants of 1.7 x 10(-11) M (high-affinity receptors) and 1.3 x 10(-9) M (low-affinity receptors). Unlabeled BDNF competed with 125I-BDNF for binding to the high-affinity receptors with an inhibition constant essentially identical to the dissociation constant of the labeled protein: 1.2 x 10(-11) M. The association and dissociation rates from both types of receptors were also determined, and the dissociation constants calculated from these kinetic experiments were found to correspond to the results obtained from steady-state binding. The number of high-affinity receptors (a few hundred per cell soma) was 15 times lower than that of low-affinity receptors. No high-affinity receptors were found on sympathetic neurons, known not to respond to BDNF, although specific binding of 125I-BDNF to these cells was detected at a high concentration of the radioligand. These results are discussed and compared with those obtained with nerve growth factor on the same neuronal populations.
Rodriguez-Tébar
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A
Max-Planck-Institute for Psychiatry, Department of Neurochemistry, Martinsried, Federal Republic of Germany.
Barde
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YA
D000818:Animals; D019208:Brain-Derived Neurotrophic Factor; D002642:Chick Embryo; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism; D006706:Homeostasis; D007457:Iodine Radioisotopes; D007700:Kinetics; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D013564:Sympathetic Nervous System / Q000166:cytology / Q000378:metabolism
D019208:Brain-Derived Neurotrophic Factor; D007457:Iodine Radioisotopes; D009419:Nerve Tissue Proteins; D011956:Receptors, Cell Surface