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Sci. Signal., 9 August 2011
Vol. 4, Issue 185, p. ra51
[DOI: 10.1126/scisignal.2001707]


Distinct Phosphorylation Sites on the β2-Adrenergic Receptor Establish a Barcode That Encodes Differential Functions of β-Arrestin

Kelly N. Nobles1*, Kunhong Xiao2*{dagger}, Seungkirl Ahn2, Arun K. Shukla2,3, Christopher M. Lam2, Sudarshan Rajagopal2, Ryan T. Strachan2, Teng-Yi Huang2, Erin A. Bressler2, Makoto R. Hara2, Sudha K. Shenoy2,4, Steven P. Gygi5, and Robert J. Lefkowitz1,2,3{dagger}

1 Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.
2 Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
3 Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710, USA.
4 Department of Cell Biology, Duke University Medical Center, Durham, NC 27710, USA.
5 Department of Cell Biology, Harvard University Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.

* These authors contributed equally to this work.

Abstract: Phosphorylation of G protein–coupled receptors (GPCRs, which are also known as seven-transmembrane spanning receptors) by GPCR kinases (GRKs) plays essential roles in the regulation of receptor function by promoting interactions of the receptors with β-arrestins. These multifunctional adaptor proteins desensitize GPCRs, by reducing receptor coupling to G proteins and facilitating receptor internalization, and mediate GPCR signaling through β-arrestin–specific pathways. Detailed mapping of the phosphorylation sites on GPCRs targeted by individual GRKs and an understanding of how these sites regulate the specific functional consequences of β-arrestin engagement may aid in the discovery of therapeutic agents targeting individual β-arrestin functions. The β2-adrenergic receptor (β2AR) has many serine and threonine residues in the carboxyl-terminal tail and the intracellular loops, which are potential sites of phosphorylation. We monitored the phosphorylation of the β2AR at specific sites upon stimulation with an agonist that promotes signaling by both G protein–mediated and β-arrestin–mediated pathways or with a biased ligand that promotes signaling only through β-arrestin–mediated events in the presence of the full complement of GRKs or when either GRK2 or GRK6 was depleted. We correlated the specific and distinct patterns of receptor phosphorylation by individual GRKs with the functions of β-arrestins and propose that the distinct phosphorylation patterns established by different GRKs establish a "barcode" that imparts distinct conformations to the recruited β-arrestin, thus regulating its functional activities.

{dagger} To whom correspondence should be addressed. E-mail: lefko001{at} (R.J.L.); khxiao{at} (K.X.)

Citation: K. N. Nobles, K. Xiao, S. Ahn, A. K. Shukla, C. M. Lam, S. Rajagopal, R. T. Strachan, T.-Y. Huang, E. A. Bressler, M. R. Hara, S. K. Shenoy, S. P. Gygi, R. J. Lefkowitz, Distinct Phosphorylation Sites on the β2-Adrenergic Receptor Establish a Barcode That Encodes Differential Functions of β-Arrestin. Sci. Signal. 4, ra51 (2011).

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