+ Translate
+ Most Popular
Gaucher's disease;thirty-two years experience at Siriraj Hospital
A study of Macrobathra Meyrick from China (Lepidoptera, Cosmopterigidae)
First occurrence in ores of tetragonal chalcocite
Effects of trace element nutrition on sleep patterns in adult women
N.Z. range management guidelines. 2. Design of grazing management systems for tussock country
A case of lipoma of the esophagus
A revision of world Acanthosomatidae (Heteroptera: Pentatomidae): keys to and descriptions of subfamilies, tribes and genera, with designation of types
Life history of the coronate scyphozoan Linuche unguiculata (Swartz, 1788)
Perceptual restoration of obliterated sounds
Mutagenicity studies on two chromium(III) coordination compounds
The formation of the skeleton. I. Growth of a long bone. 1st appearance of a center of calcification
Leucopenia and abnormal liver function in travellers on malaria chemoprophylaxis
The joint commission: four key root causes loom large in sentinel event data
Treatment of vitiligo with topical 15% lactic acid solution in combination with ultra violet-A
Behaviour of dairy cows within three hours after feed supply: I. Influence of housing type and time elapsing after feed supply
Observations of the propagation velocity and formation mechanism of burst fractures caused by gunshot
Management and control of patients with type 2 diabetes mellitus in Lebanon: results from the International Diabetes Management Practices Study (IDMPS)
The diet composition and nutritional knowledge of patients with anorexia nervosa
Physoporella croatica Herak, 1958 of the Slovak karst Anisian (Slovakia, the West Carpathians Mts.)
Bright lights, big noise. How effective are vehicle warning systems?
Ein Plesiosaurier-Rest mit Magensteinen aus mittlerem Lias von Quedlinburg
Incidence of Chlamydia trachomatis in patients with sterility
Monster soup: the microscope and Victorian fantasy
Preliminary tests with residual sprays against poultry lice
Duration of the life of plants in phylogeny

Purification of the neurotensin receptor from mouse brain by affinity chromatography

Purification of the neurotensin receptor from mouse brain by affinity chromatography

Journal of Biological Chemistry 264(10): 5559-5563

ISSN/ISBN: 0021-9258

PMID: 2538423

The neurotensin receptor was purified from newborn mouse brain by affinity chromatography. Active neurotensin binding sites were solubilized from brain homogenates using the nondenaturing detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (CHAPS) in the presence of cholesteryl hemisuccinate. Chromatography of the soluble extract on SP-Sephadex C-25 and hydroxylapatite eliminated 50% of proteins without loss of neurotensin binding activity. This prepurified material was loaded into an affinity column prepared by coupling neurotensin (2-13) to glutaraldehyde-activated Ultrogel AcA22. Nonspecifically adsorbed proteins were eliminated by extensive washing, and the receptor was eluted with a buffer containing 1 M NaCl, 0.1% CHAPS, and 0.02% cholesteryl hemisuccinate. After desalting, the purified receptor bound 125I-labeled neurotensin with a dissociation constant of 0.26 nM and retained its specificity towards a series of neurotensin analogues. The desalted NaCl eluate appeared on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a major band of molecular weight 100,000 which was identified as the receptor by affinity labeling with 125I-labeled neurotensin in the presence of disuccinimidyl suberate. The purity of the mouse brain receptor eluted from the affinity column was estimated to be 78%. Electroelution of the 100-kDa protein band gave an homogenous preparation of receptor. Very similar results were obtained with CHAPS-solubilized neurotensin receptors from rat and rabbit brain.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 007710343

Download citation: RISBibTeXText

Related references

Purification of neurotensin receptor from newborn mouse brain by affinity chromatography. Conn, P M (Ed ) Methods in Neurosciences, Vol 5 Neuropeptide Technology: Gene Expression And Neuropeptide Receptors Xvii+570p Academic Press, Inc : San Diego, California, Usa; London, England, Uk Illus 203-215, 1991

Quantitative evaluation of neurotensin receptor purification by immobilized metal affinity chromatography. Protein Expression and Purification 11(1): 53-60, 1997

Solubilization and purification of a high affinity neurotensin receptor from newborn human brain. Brain Research 639(2): 245-252, 1994

Purification of the IL-2 receptor (TAC) by ligand-affinity chromatography and utilization of the immobilized receptor for receptor-affinity chromatography (RAC) purification of IL-2, mutant IL-2, and IL-2 fusion proteins. Journal of Investigative Dermatology 94(6 Suppl): 158S-163S, 1990

Purification of active neurotensin receptor from mouse brain. Aubry, A , M Marraud And B Vitoux (Ed ) Colloque Inserm (Institut National De La Sante Et De La Recherche Medicale), Vol 174 Deuxieme Forum Peptides; (Inserm (National Institute Of Health And Medical Research) Colloquium), Vol 174 Second Forum on Peptides; Nancy, France, May 2-6, 1988 Xxxix+612p Les Editions Inserm: Paris, France; John Libbey And Co Ltd : London, England, Uk Illus Paper 163-166, 1989

Regional and cellular distribution of low affinity neurotensin receptor mRNA in adult and developing mouse brain. Journal of Comparative Neurology 394(3): 344-356, 1998

Purification and characterization of the mu opiate receptor from rat brain using affinity chromatography. Proceedings of the National Academy of Sciences of the United States of America 82(2): 594-598, 1985

Purification of a 1 adenosine receptor of rat brain membranes by affinity chromatography. Jacobson, K A , J W Daly And V Manganiello (Ed ) Purines in Cellular Signaling: Targets For New Drugs; Conference, Rockville, Maryland, Usa, September 17-20, 1989 Xv+448p Springer-Verlag New York, Inc : Secaucus, New Jersey, Usa; Berlin, West Germany Illus 65-68, 1990

Purification of a rat brain nicotinic receptor by affinity and immunoaffinity chromatography. Rand, M J And K Thurau (Ed ) Icsu (International Council Of Scientific Unions) Press Symposium Series, Vol 9 The Pharmacology Of Nicotine; Satellite Symposium Of The Tenth International Congress Of Pharmacology, Gold Coast, Queensland, Australia, September 4-6, 1987 Xii+418p Irl Press Ltd : Oxford, England, Uk; Washington, D C , Usa Illus Paper 248-250, 1988

Rapid purification of protein phosphatase 2A from mouse brain by microcystin-affinity chromatography. Febs Letters 279(1): 115-118, 1991

Purification to homogeneity of an active opioid receptor from rat brain by affinity chromatography. Proceedings of the National Academy of Sciences of the United States of America 91(10): 4574-4578, 1994

Partial purification of rat brain and testicular a 1 adenosine receptor by affinity chromatography. Society for Neuroscience Abstracts 14(1): 110, 1988

Changes in Mouse Brain Neurotensin Receptor Density Following Chronic Infusion of Neurotensin. Peptides 16(3): 501-504, 1995

Purification of gamma aminobutyric acid receptor from rat brain by benzodiazepine affinity column chromatography. Neurochemical Research 9(8): 1139-1140, 1984

Purification of gamma-aminobutyric acid (GABA) receptor from rat brain by affinity column chromatography using a new benzodiazepine, 1012-S, as an immobilized ligand. Brain Research 323(2): 219-226, 1984