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J. Biol. Chem., Vol. 277, Issue 47, 44925-44931, November 22, 2002
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From the Département de Biochimie and Groupe de Recherche sur
le Système Nerveux Autonome, Université de Montréal,
Montréal, Quebec H3C 3J7, Canada
Quantitative bioluminescence resonance energy
transfer (BRET) analysis was applied to the study of
Quantitative Assessment of
1- and
2-Adrenergic Receptor Homo- and Heterodimerization
by Bioluminescence Resonance Energy Transfer*
§,¶,
1- and 2-adrenergic receptor homo-
and heterodimerization. To assess the relative affinity between each of
the protomers, BRET saturation experiments were carried out in HEK-293T
cells. 1- and 2-adrenergic receptors were
found to have similar propensity to engage in homo- and heterotropic interactions suggesting that, at equivalent expression levels of the
two receptor subtypes, an equal proportion of homo- and heterodimers
would form. Analysis of the data also revealed that, at equimolar
expression levels of energy donor and acceptor, more than 80% of the
receptor molecules exist as dimers and that this high incidence of
receptor dimerization is insensitive to receptor density for expression
levels varying between 1.4 and 26.9 pmol of receptor/mg of membrane
protein. Taken together, these results indicate that most of the
receptors expressed in cells exist as constitutive dimers and that, at
least in undifferentiated fibroblasts, the proportion of homo- and
heterodimers between the closely related 1- and
2-adrenergic receptors is determined by their relative levels of expression.
*
This work was supported in part by grants from the Canadian
Institute for Health Research (CIHR) and the Heart and Stroke Foundation of Canada (to M. B.).The costs of publication of this article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Both authors contributed equally to this work.
§
Held a studentship from the Fonds de la Recherche en Santé du
Québec.
¶
Hold studentships from the CIHR.
Holds a Canada Research Chair in Molecular and Cellular
Pharmacology. To whom correspondence should be addressed: Dépt.
de Biochimie, Université de Montréal, C.P. 6128, Succursale
Centre-Ville, Montréal, Quebec H3C 3J7, Canada. Tel.:
514-343-6372; Fax: 514-343-2210; E-mail:
michel.bouvier@umontreal.ca.
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