Radioligand Binding Assay for the Simultaneous Determination of β1- and β2-Adrenergic Receptors in Human Blood Cells

Radioligand Binding Assay for the Simultaneous Determination of β1- and β2-Adrenergic Receptors in Human Blood Cells

In this study, we propose a method for the separate determination of both types of adrenergic receptors based on radioligand binding analysis using 125 I-iodocyanopindolol, comprising three measurements: (1) without competing ligands, (2) in the presence of selective ligand ICI 118,551 (0.25 μM), an...

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Bibliographic Details
Published in:Russian journal of bioorganic chemistry Vol. 48; no. 5; pp. 1002 - 1010
Main Authors: Shevelev, A. Y., Kashirina, N. M., Lipatova, L. N., Yanushevskaya, E. V., Peklo, M. M., Rybalkin, I. N., Rutkevich, P. N., Chusovitina, O. K., Skoblova, N. A., Skoblov, Yu. S., Vlasik, T. N., Zykov, K. A.
Format: Journal Article
Language:English
Published: Moscow Pleiades Publishing 01-10-2022
Springer Nature B.V
Subjects:
Adrenergic receptors
Binding
Biochemistry
Biomedical and Life Sciences
Biomedicine
Bioorganic Chemistry
Blood cells
Error analysis
Life Sciences
Ligands
Organic Chemistry
Receptors (physiology)
radioligand binding assay
β
and β
adrenergic receptors
human cells
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Summary:In this study, we propose a method for the separate determination of both types of adrenergic receptors based on radioligand binding analysis using 125 I-iodocyanopindolol, comprising three measurements: (1) without competing ligands, (2) in the presence of selective ligand ICI 118,551 (0.25 μM), and (3) in the presence of two selective ligands, ICI 118,551 and CGP 20712 (0.25 μM each). The technique was tested on a model system of two transgenic cell lines ADL-7A and A2R9 with the expression of recombinant β 1 - and β 2 -adrenergic receptors. If the ratio of the number of β 1 -adrenergic receptors to β 2 -adrenergic receptors is 1 : 10, the measurement error is about 15%. Analysis of nine cell lines representing different types of blood cells showed the presence of β 2 -adrenergic receptors in Daudi, Raji, Dami, K-562, HL-60, U-937, and THP-1 cells and their absence in Jurkat and MOLT-4 cells. β 1 -Adrenergic receptors are reliably recorded only in THP-1 cells of monocytic origin. In other cell lines, with the exception of Dami, the number of β 1 -adrenergic receptors was found below the detection limit, estimated as 250 molecules per cell. Measurements performed on the peripheral blood mononuclear cells of seven healthy donors showed the presence of β 2 -adrenergic receptors in the range from 1000 to 2500 molecules per cell, while the content of β 1 -adrenergic receptors in all cases appeared to be on the border or beyond the detection limit.
ISSN:1068-1620
1608-330X
DOI:10.1134/S1068162022050211