Solubilization of nerve growth factor receptors of rabbit superior cervical ganglia

S. P. Banerjee, P. Cuatrecasas, S. H. Snyder

Research output: Contribution to journalArticlepeer-review

Abstract

Nerve growth factor (NGF) receptors of rabbit superior cervical ganglia can be solubilized by treatment with detergents and readily assayed in the soluble state. Triton X 100 and deoxycholate reduce specific binding of NGF to ganglia membranes. In membranes treated with Triton X 100 (0.5 to 2.0%) the reduction in NGF binding by membranes is accompanied by a corresponding increase in binding in the supernatant fluid. NGF binding in soluble preparations can be rapidly assayed by precipitating NGF bound to receptors with polyethylene glycol under conditions in which unbound NGF is not precipitated. NGF binding to soluble preparations is saturable whether evaluated by the binding of 125I NGF or by diluting 125I NGF with native NGF. Using both techniques, the dissociation constant for NGF binding to soluble receptors is about 0.2 nM, the same as its dissociation constant from receptor sites in intact membranes. NGF binding to soluble receptors displays a high degree of peptide specificity, similar to receptor sites in intact membranes of superior cervical ganglia. A method of labeling NGF with 125I 3(4 hydroxyphenyl) propionic acid N hydroxysuccinimide ester is described which leads to binding properties that are superior to those obtained with previously described 125I NGF preparations.

Original languageEnglish (US)
Pages (from-to)5680-5685
Number of pages6
JournalJournal of Biological Chemistry
Volume251
Issue number18
StatePublished - 1976

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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