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  Synthesis and properties of fluorescent ß-adrenoceptor ligands

Heithier, H., Hallmann, D., Boege, F., Reiländer, H., Dees, C., Jaeggi, K. D., et al. (1994). Synthesis and properties of fluorescent ß-adrenoceptor ligands. Biochemistry, 33(31), 9126-9134. doi:10.1021/bi00197a015.

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 Creators:
Heithier, Helmut1, Author
Hallmann, Dieter2, Author           
Boege, Fritz3, Author
Reiländer, Helmut4, Author           
Dees, Christian1, Author
Jaeggi, Knut D.5, Author
Arndt-Jovin, Donna J.6, Author           
Jovin, Thomas M.6, Author           
Helmreich, Ernst J. M.1, Author
Affiliations:
1Medizinische Universitätsklinik, Abteilung für Klinische Biochemie und Pathobiochemie, Würzburg, Germany, ou_persistent22              
2Department of Membrane Biophysics, MPI for biophysical chemistry, Max Planck Society, ou_578579              
3Medizinische Poliklinik der Universität, 97070 Würzburg, Germany,, ou_persistent22              
4Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068290              
5Ciba-Geigy Ltd., CH-4002 Basel, Switzerland, ou_persistent22              
6Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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 Abstract: We describe the synthesis of bordifluoropyrromethene (BODIPY), fluorescein, and related fluorescent derivatives of the beta-adrenergic ligand CGP 12177. With these probes we screened insect (Sf9) cells stably transformed with the human ß2-adrenoceptor gene and expressing (2-3.5) x 105 human ß2-adrenoceptors per cell. Among these derivatives only BODIPY-CGP gave a receptor-specific signal sufficiently strong for measuring the on- and off-rate constants and the equilibrium dissociation constant of beta-adrenoceptor-specific binding by spectrofluorometry or photon counting. Similar KD values for BODIPY-CGP binding were obtained by kinetic measurements (approx. 250 pM) and under equilibrium conditions (400 +/- 180 pM), and these were in the same range as those obtained with [3H]CGP 12177 (200 +/- 32 pM). The cell-bound fluorescence could be quenched specifically with nonfluorescent CGP 12177 to near background levels. The disposition of the ß2-adrenoceptors in BODIPY-CGP-stained Sf9 cells was mainly restricted to the cell surface at 4 and 30 degrees C. Hence, beta-adrenoceptor-expressing cells can be stained specifically with BODIPY-CGP, and beta-adrenoceptors on a single cell can be assessed by photon counting under the fluorescence microscope. Cells can also be scanned by fluorescence-activated flow cytometry.

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Language(s): eng - English
 Dates: 1994-05-041994-01-272002-05-011994-08-09
 Publication Status: Issued
 Pages: 9
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1021/bi00197a015
PMID: 7914090
 Degree: -

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Title: Biochemistry
Source Genre: Journal
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Publ. Info: Columbus, Ohio : American Chemical Society
Pages: - Volume / Issue: 33 (31) Sequence Number: - Start / End Page: 9126 - 9134 Identifier: ISSN: 0006-2960
CoNE: https://pure.mpg.de/cone/journals/resource/954925384103