Nerve growth factor receptor from rabbit sympathetic ganglia membranes. Relationship between subforms.
The receptor for nerve growth factor (NGF) was purified from Triton X-100 extracts of sympathetic ganglia membranes by affinity chromatography on NGF-Sepharose. Elution of purified receptor was accomplished at pH 5 in the presence of 1 M NaCl. Sodium dodecyl sulfate gel electrophoresis of the purified iodinated receptor showed three major bands at Mr = 126,000, Mr = 105,000, and Mr = 81,000. Affinity labeling of the purified receptor using 125I-NGF and the photoreactive agent N-hydroxysuccinimidyl-p-azidobenzoate resulted in two major cross-linked complexes corresponding to Mr = 135,000 and Mr = 110,000. This labeling pattern is similar to that observed with sympathetic ganglia membranes (Massague, J., Guillette, B. J., Czech, M. P., Morgan, C. J., and Bradshaw, R. A. (1981) J. Biol. Chem. 256, 9419-9424) and indicates that these two forms do not arise from the cross-linking procedure. Reaction of the photoaffinity labeled NGF receptors with increasing amounts of trypsin resulted in a progressive decrease in the high molecular weight complex with a concomitant increase in the low molecular weight form. When the larger complex was isolated by electroelution from a sodium dodecyl sulfate gel and treated with trypsin, a species corresponding to Mr = 100,000 was generated. These observations are best explained by a precursor-product relationship for the two NGF receptor species of sympathetic neurons.
D000345:Affinity Labels; D000818:Animals; D005728:Ganglia, Sympathetic / Q000032:analysis*; D007457:Iodine Radioisotopes; D008297:Male; D051379:Mice; D008970:Molecular Weight; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000032:analysis / Q000302:isolation & purification*; D017475:Receptors, Nerve Growth Factor; D014357:Trypsin / Q000494:pharmacology
D000345:Affinity Labels; D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D017475:Receptors, Nerve Growth Factor; D011092:Polyethylene Glycols; D017830:Octoxynol; D014357:Trypsin