Hepatic α-adrenergic receptors. Identification and subcellular localization using [3H]dihydroergocryptine

W. R. Clarke, Larry Jones, R. J. Lefkowitz

Research output: Contribution to journalArticle

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Abstract

Recently, several workers have shown that adrenergic control of hepatic carbohydrate metabolism has the characteristics of an α-receptor-mediated process. Using the rat liver membrane preparation of Neville (Neville, D, (1968) Biochim. Biophys. Acta 154, 540-552), α-adrenergic receptors have been identified using the ligand [3H]dihydroergocryptine. The receptors are saturable and of high affinity. Scatchard analysis yields a K(D) of 1.8 nM with 1.7 ± 0.55 pmol of sites/mg of protein. Competition of dihydroergocryptine binding with various pharmacologic agents yields the typical (α-adrenergic potency series: (-)-epinephrine > (-)-norepinephrine > (-)-isoproterenol. (-)-Isomers are more potent than (+)-isomers. The α-blocker phentolamine is 3.4 orders of magnitude more potent than the β-blocker propranolol. To determine subcellular localization of α-adrenergic receptors, livers were fractionated into a crude homogenate, a 1500 x g pellet, and the purified membrane preparation used previously for binding. Specific dihydroergocryptine binding, ouabain-inhibitable (Na,K)-ATPase, and F--stimulated adenylate cyclase activities, were followed in these fractions. Specific binding was enriched, relative to that in the crude homogenate, 2.88-fold in the pellet and 6.28-fold in the membranes. Similarly, (Na,K)-ATPase activity was enriched 2.6-fold in the pellet and 7.1-fold in the membranes while adenylate cyclase activity was enriched 2.9-fold in the pellet and 3.5-fold in the membranes. It is concluded that hepatic α-adrenergic receptors are likely concentrated in the plasma membranes.

Original languageEnglish (US)
Pages (from-to)5975-5979
Number of pages5
JournalJournal of Biological Chemistry
Volume253
Issue number17
StatePublished - 1978
Externally publishedYes

Fingerprint

Dihydroergocryptine
Adrenergic Receptors
Membranes
Liver
Adenylyl Cyclases
Isomers
Adrenergic Agents
Adenosine Triphosphatases
Phentolamine
Carbohydrate Metabolism
Ouabain
Cell membranes
Isoproterenol
Propranolol
Epinephrine
Rats
Norepinephrine
Cell Membrane
Ligands

ASJC Scopus subject areas

  • Biochemistry

Cite this

Hepatic α-adrenergic receptors. Identification and subcellular localization using [3H]dihydroergocryptine. / Clarke, W. R.; Jones, Larry; Lefkowitz, R. J.

In: Journal of Biological Chemistry, Vol. 253, No. 17, 1978, p. 5975-5979.

Research output: Contribution to journalArticle

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