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Solubilization and characterization of active neurotensin receptors from mouse brain

Journal Article · · J. Biol. Chem.; (United States)
OSTI ID:6924816
; ; ;
Neurotensin receptors were solubilized from mouse brain using the zwitterionic detergent 3-((3-cholamidopropyl)dimethylammonio)-1-propanesulfonic acid (CHAPS). The binding of /sup 125/I-labeled (Tyr/sup 3/)neurotensin to the soluble fraction was time-dependent, saturable, and reversible. Unlabeled neurotensin and its analogues acetylneurotensin (8-13), neurotensin (9-13), and neurotensin (1-12) competitively antagonized the binding of /sup 125/I-labeled (Tyr/sup 3/)neurotensin to CHAPS-solubilized extracts with relative potencies similar to those observed with membrane-bound receptors. Scatchard analysis of equilibrium binding data indicated that the soluble extract contained a single class of neurotensin binding sites with a K/sub d/ of 0.36 nM and a B/sub m/ of 63 fmol/mg. As already observed with membrane-bound receptors, the affinity of neurotensin for the soluble binding activity was decreased by Na/sup +/ ions. By contrast, soluble receptors were no longer sensitive to GTP and the antihistamine drug levocabastine. A molecular weight of about 100,000 was determined for soluble neurotensin receptors both under native conditions by gel filtration on Ultrogel AcA 34 and under denaturating conditions by sodium dodecyl sulfate-polyacrylamide gel electrophoresis after photoaffinity labeling.
Research Organization:
Universite de Nice (France)
OSTI ID:
6924816
Journal Information:
J. Biol. Chem.; (United States), Journal Name: J. Biol. Chem.; (United States) Vol. 263:1; ISSN JBCHA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
BRAIN
CENTRAL NERVOUS SYSTEM
CONFIGURATION INTERACTION
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
KINETICS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
MICE
MOLECULAR STRUCTURE
MOLECULAR WEIGHT
NERVOUS SYSTEM
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PEPTIDES
POLYPEPTIDES
PROTEINS
PURIFICATION
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
RODENTS
VERTEBRATES