title: Purification of the neurotensin receptor from bovine brain.
abstract: 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 (Kd = 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 Mr 72,000. Under nonreducing conditions the apparent Mr, however, was 50,000, suggesting the presence of intramolecular disulfide bonds. The purified neurotensin receptor was judged to be homogeneous, 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.
authors.affiliation: Medical Research Council Molecular Neurobiology Unit, Medical Research Council Centre, Cambridge, Great Britain.
authors.forename: A
authors.initials: A
authors.lastname: Mills
authors.forename: C D
authors.initials: CD
authors.lastname: Demoliou-Mason
authors.forename: E A
authors.initials: EA
authors.lastname: Barnard
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D002846:Chromatography, Affinity; D007700:Kinetics; D008970:Molecular Weight; D009496:Neurotensin / Q000378:metabolism*; D018028:Receptors, Neurotensin; D017981:Receptors, Neurotransmitter / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D018028:Receptors, Neurotensin; D017981:Receptors, Neurotransmitter; D009496:Neurotensin