Solubilization and characterization of active neurotensin receptors from mouse brain
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), Vol. 263:1
- Country of Publication:
- United States
- Language:
- English
Similar Records
Murine interleukin 1 receptor. Direct identification by ligand blotting and purification to homogeneity of an interleukin 1-binding glycoprotein
Characterization of solubilized human and rat brain US -endorphin-receptor complex
Related Subjects
POLYPEPTIDES
BIOCHEMICAL REACTION KINETICS
CONFIGURATION INTERACTION
RECEPTORS
MOLECULAR STRUCTURE
PURIFICATION
BRAIN
IODINE 125
LABELLED COMPOUNDS
MICE
MOLECULAR WEIGHT
ANIMALS
BETA DECAY RADIOISOTOPES
BODY
CENTRAL NERVOUS SYSTEM
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPES
KINETICS
MAMMALS
MEMBRANE PROTEINS
NERVOUS SYSTEM
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PEPTIDES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RODENTS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques