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J. Biol. Chem., Vol. 268, Issue 5, 3201-3208, 02, 1993
S Pippig, S Andexinger, K Daniel, M Puzicha, MG Caron, RJ Lefkowitz and MJ Lohse
Receptor-specific or homologous desensitization of beta 2-adrenergic
receptors is thought to be effected via phosphorylation of the receptor by
the beta-adrenergic receptor kinase (beta ARK), followed by binding of
beta-arrestin. We have generated stably transfected Chinese hamster ovary
cell lines overexpressing either of the two regulatory proteins and also
expressing low or high levels of beta 2-adrenergic receptors (approximately
80 and approximately 600 fmol/mg of membrane protein). In these cells, we
studied the process of desensitization induced by the beta-adrenergic
receptor agonist isoproterenol. In cells expressing high levels of beta
2-adrenergic receptors, desensitization to high concentrations of
isoproterenol (previously shown to be mediated by both beta ARK and protein
kinase A) amounted to approximately 50% in control cells, approximately 80%
in beta ARK-overexpressing cells, and approximately 90% in
beta-arrestin-overexpressing cells. In cells expressing low levels of beta
2-adrenergic receptors, these values were approximately 50, approximately
60, and approximately 60%, respectively. Desensitization to low
concentrations of isoproterenol (previously shown to be essentially protein
kinase A-mediated and not receptor-specific, i.e. heterologous) was not
affected by overexpression of either beta ARK or beta-arrestin. These data
suggest that in cells expressing high levels of beta 2-adrenergic
receptors, beta-arrestin and beta ARK become limiting for homologous
receptor desensitization. They provide further support for the involvement
of these two proteins in the regulation of beta 2-adrenergic receptor
function.
Overexpression of beta-arrestin and beta-adrenergic receptor kinase augment desensitization of beta 2-adrenergic receptors
Laboratory of Molecular Biology, University of Munich, Max-Planck- Institute of Biochemistry, Martinsried, Federal Republic of Germany.
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