Purification of the neurotensin receptor from bovine brain

Journal Article · Tue Jan 05 00:00:00 EST 1988 · J. Biol. Chem.; (United States)

OSTI ID:6838007

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

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-/sup 3/H) neurotensin with an apparent affinity (K/sub 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/sub r/ 72,000. Under nonreducing conditions the apparent M/sub 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.

• Research Organization: Medical Research Council Centre, Cambridge (England)
• OSTI ID: 6838007
• Journal Information: J. Biol. Chem.; (United States), Vol. 263:1
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
RECEPTORS
BIOCHEMICAL REACTION KINETICS
PURIFICATION
BRAIN
CATTLE
ELECTROPHORESIS
LIGANDS
PEPTIDES
TRITIUM COMPOUNDS
ANIMALS
BODY
CENTRAL NERVOUS SYSTEM
DOMESTIC ANIMALS
KINETICS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
NERVOUS SYSTEM
ORGANIC COMPOUNDS
ORGANS
PROTEINS
REACTION KINETICS
RUMINANTS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques