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Regulation of cell signaling: GRK2 dependent phosphorylation of alpha2A‐adrenergic receptors (802.12)
Author(s) -
Hebert Aaron,
SterneMarr Rachel
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.802.12
Subject(s) - phosphorylation , beta adrenergic receptor kinase , kinase , microbiology and biotechnology , receptor , g protein coupled receptor kinase , chemistry , signal transduction , biology , g protein coupled receptor , biochemistry
Cells relay information, such as the fight or flight response, via chemicals, such as adrenalin, and one of two classes of adrenergic receptors, α and β. The α2A‐adrenergic receptor (α2AAR), has been shown to play a key role in vasoconstriction in some parts of the cardiovascular system, and is the target of pharmaceuticals. Proper regulation of cellular signals by desensitization of the receptor is essential to maintaining the organismal status quo or homeostasis. Phosphorylation, or the addition of a highly charged phosphate group, is carried out by kinases and is a common mechanism of receptor desensitization. We study a kinase, called GRK2, that phosphorylates adrenergic receptors after they are treated with adrenalin. One way to study the phosphorylation of the α2AAR in intact cells is to use antibodies that specifically recognize the phosphorylated receptor. Because no commercially available α2AAR phosphosite antibodies currently exist, we set out to generate such a reagent. A functional phosphosite antibody that detects α2AAR phosphorylation was isolated and characterized. This antibody that we call Rb617 was used to test whether the phosphorylation of α2AAR in intact cells was adrenalin‐induced and GRK2‐dependent in primate cells in culture. We found that the α2AAR was phosphorylated in the absence of adrenalin and exogenous GRK2 suggesting that, in addition to GRK2, other cellular kinases modify the α2AAR. Further intact cell phosphorylation assays will likely include the use of inhibitors to minimize the effect of phosphorylation due to these other kinases so that we might study phosphorylation by GRK2.