Affinity Chromatography of the D1Dopamine Receptor from Rat Corpus Striatum

Jay A. Gingrich, Nourdine Amlaiky, Susan Senogles, Wei K. Chang, Robert D. Mcquade, Joel G. Berger, Marc G. Caron

Research output: Contribution to journalArticle

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Abstract

The D1dopamine receptor from rat corpus striatum has been purified 200-250-fold by using a newly developed biospecific affinity chromatography matrix based on a derivative of the D1selective antagonist SCH 23390. This compound, (RS)-5-(4-aminophenyl)-8-chloro-2,3,4,5-tetrahydro-3-methyl-l/f-3-benzazepin-7-ol (SCH 39111), possesses high affinity for the D1receptor and, when immobilized on Sepharose 6B through an extended spacer arm, was able to adsorb digitonin-solubilized D1receptors. The interaction between the solubilized receptor and the affinity matrix was biospecific. Adsorption of receptor activity could be blocked in a stereoselective fashion [SCH 23390 > SCH 23388; (+)-butaclamol > (-)-butaclamol]. The elution of [3H]SCH 23390 activity from the gel demonstrated similar stereoselectivity for antagonist ligands. Agonists eluted receptor activity with a rank order of potency consistent with that of a D1receptor [apomorphine >; dopamine > (-)-epinephrine ≫ LY 171555 > serotonin]. SCH 39111-Sepharose absorbed 75-85% of the soluble receptor activity, and after the gel was washed extensively, 35-55% of the absorbed receptor activity could be eluted with 100 μM (+)-butaclamol with specific activities ranging from 250 to 450 pmol/mg of protein. The affinity-purified receptor retains the ligand binding characteristics of a D1dopamine receptor. This affinity chromatography procedure should prove valuable in the isolation and molecular characterization of the D1dopamine receptor.

Original languageEnglish (US)
Pages (from-to)3907-3912
Number of pages6
JournalBiochemistry
Volume27
Issue number11
DOIs
StatePublished - May 1 1988

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Butaclamol
Affinity chromatography
Corpus Striatum
Affinity Chromatography
Rats
Sepharose
4,4a,5,6,7,8,8a,9-octahydro-5-propyl-1H-pyrzolo(3,4-g)quinoline
Gels
Ligands
Stereoselectivity
Digitonin
Apomorphine
Epinephrine
Adsorption
Dopamine
Serotonin
Derivatives
SCH 23390
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Gingrich, J. A., Amlaiky, N., Senogles, S., Chang, W. K., Mcquade, R. D., Berger, J. G., & Caron, M. G. (1988). Affinity Chromatography of the D1Dopamine Receptor from Rat Corpus Striatum. Biochemistry, 27(11), 3907-3912. https://doi.org/10.1021/bi00411a003

Affinity Chromatography of the D1Dopamine Receptor from Rat Corpus Striatum. / Gingrich, Jay A.; Amlaiky, Nourdine; Senogles, Susan; Chang, Wei K.; Mcquade, Robert D.; Berger, Joel G.; Caron, Marc G.

In: Biochemistry, Vol. 27, No. 11, 01.05.1988, p. 3907-3912.

Research output: Contribution to journalArticle

Gingrich, JA, Amlaiky, N, Senogles, S, Chang, WK, Mcquade, RD, Berger, JG & Caron, MG 1988, 'Affinity Chromatography of the D1Dopamine Receptor from Rat Corpus Striatum', Biochemistry, vol. 27, no. 11, pp. 3907-3912. https://doi.org/10.1021/bi00411a003
Gingrich JA, Amlaiky N, Senogles S, Chang WK, Mcquade RD, Berger JG et al. Affinity Chromatography of the D1Dopamine Receptor from Rat Corpus Striatum. Biochemistry. 1988 May 1;27(11):3907-3912. https://doi.org/10.1021/bi00411a003
Gingrich, Jay A. ; Amlaiky, Nourdine ; Senogles, Susan ; Chang, Wei K. ; Mcquade, Robert D. ; Berger, Joel G. ; Caron, Marc G. / Affinity Chromatography of the D1Dopamine Receptor from Rat Corpus Striatum. In: Biochemistry. 1988 ; Vol. 27, No. 11. pp. 3907-3912.
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