Purification of the neurotensin receptor from bovine brain

A. Mills, C. D. Demoliou-Mason, E. A. Barnard

Research output: Contribution to journalArticlepeer-review


The neurotensin receptor protein, solubilized with digitonin/asolectin from bovine cerebral cortex membranes, was purified to apparent homogeneity by affinity chromatography using immobilized neurotensin. The product exhibits saturable and specific binding of [3,11-tyrosyl-3,5- 3H]neurotensin with an apparent affinity (K(d) = 5.5 nM) comparable to that measured in intact membranes and crude soluble extracts. The affinity-purified material, after reduction with 100 mM dithiothreitol, in denaturing gel electrophoresis showed a single polypeptide of M(r) 72,000. Under non-reducing conditions the apparent M(r), however, was 50,000, suggesting the presence of intramolecular disulfide bonds. The purified neurotensin receptor was judged to be homogenous, in that (i) only a single polypeptide was detectable; and (ii) the overall purification was 30,000-50,000-fold, giving a specific neurotensin-binding activity close to the theoretical maximum.

Original languageEnglish
Pages (from-to)13-16
Number of pages4
JournalJournal of Biological Chemistry
Issue number1
Publication statusPublished - 1988


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