TY - JOUR
T1 - Purification of the neurotensin receptor from bovine brain
AU - Mills, A.
AU - Demoliou-Mason, C. D.
AU - Barnard, E. A.
PY - 1988
Y1 - 1988
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0023930787&partnerID=8YFLogxK
M3 - Article
C2 - 2826433
AN - SCOPUS:0023930787
SN - 0021-9258
VL - 263
SP - 13
EP - 16
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 1
ER -