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Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 305, Issue 1, pp 41–50 | Cite as

Identification and quantification of β-adrenoceptor sites in red blood cells from rats

  • G. Kaiser
  • G. Wiemer
  • G. Kremer
  • J. Dietz
  • D. Palm
Article

Summary

For the direct characterization of β-adrenoceptors in membrane preparations from the reticulocyte rich blood of rats treated with acetyl phenylhydrazide the β-adrenergic antagonist ligand (3H)(−)dihydroalprenolol (DHAP) was used.

  1. 1.

    Specific binding of DHAP demonstrated one type of binding site in these membrane preparations. There was no evidence for negative co-operativity between the sites. A mean K D -value amounting to 6.51\+-0.8 nM (n=15) was calculated from equilibrium experiments; a similar K D -value (8.25 nM) was evaluated from the ratio of the rate constants of the dissociation and association reactions. In preparations of reticulocyte rich blood (53% reticulocytes) a mean density of DHAP binding sites of 0.602\+-0.05 pmoles/mg protein (n=15) was determined. The respective value in membrane preparations from reticulocyte poor blood, i.e. from untreated animals (2% reticulocytes), amounted to only 0.224\+-0.03 pmoles/mg protein (n=5) whereas the mean K D -value remained unaltered (K D =6.84\+-2.2 nM; n=5).

     
  2. 2.

    Specific binding sites for DHAP in membranes from reticulocyte rich blood can be looked at as true \gb-adrenoceptors:

    Specific binding of DHAP was competitively inhibited by β-adrenoceptor agonists and antagonists according to the structural specificity and stereospecificity of these compounds. The K D -values for agonists increased in the order isoprenaline < adrenaline < noradrenaline < phenylephrine, the (−)enantiomers being significantly more potent than the respective (+)enantiomers. The same was true also for β-adrenoceptor antagonists.

     
  3. 3.

    Using the highly purified enantiomers of fenoterol, a compound with 2 asymmetric centres, it was demonstrated that the 1R/1\t'R isomer was highly active while the enantiomer 1S/1\t'S did not show any affinity for \gb-adrenoceptors.

     
  4. 4.

    When the K D -values evaluated from binding experiments were compared to the respective K a - and K i -values obtained from adenyl cyclase assays in the same membrane preparations there resulted direct correlations (K D \~-K a or K i ). The identity of these values for \gb-adrenoceptor agonists is in favour of an optimal coupling state of the \gb-adrenoceptor-adenyl cyclase-system in immature red blood cells from rats.

     

Key words

Red Blood Cells Rats β-Adrenoceptors Adenyl cyclase (3H) (−)Dihydroalprenolol 

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Copyright information

© Springer-Verlag 1978

Authors and Affiliations

  • G. Kaiser
    • 1
  • G. Wiemer
    • 1
  • G. Kremer
    • 1
  • J. Dietz
    • 1
  • D. Palm
    • 1
  1. 1.Zentrum der PharmakologieKlinikum der Johann Wolfgang Goethe-UniversitätFrankfurt am Main 70Federal Republic of Germany

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