Local membrane charge regulates β<sub>2</sub> adrenergic receptor coupling to G<sub>i3</sub>.

Strohman, M J; Maeda, S; Hilger, D; Masureel, M; Du, Y; Kobilka, B K · Nat Commun · 2019

basic_science · Level V

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Abstract

The β<sub>2</sub> adrenergic receptor (β<sub>2</sub>AR) signals through both G<sub>s</sub> and G<sub>i</sub> in cardiac myocytes, and the G<sub>i</sub> pathway counteracts the G<sub>s</sub> pathway. However, G<sub>i</sub> coupling is much less efficient than G<sub>s</sub> coupling in most cell-based and biochemical assays, making it difficult to study β<sub>2</sub>AR-G<sub>i</sub> interactions. Here we investigate the role of phospholipid composition on G<sub>s</sub> and G<sub>i</sub> coupling. While negatively charged phospholipids are known to enhance agonist affinity and stabilize an active state of the β<sub>2</sub>AR, we find that they impair coupling to G<sub>i3</sub> and facilitate coupling to G<sub>s</sub>. Positively charged Ca<sup>2+</sup> and Mg<sup>2+</sup>, known to interact with the negative charge on phospholipids, facilitates G<sub>i3</sub> coupling. Mutational analysis suggests that Ca<sup>2+</sup> coordinates an interaction between phospholipid and the negatively charged EDGE motif on the amino terminal helix of G<sub>i3</sub>. Taken together, our observations suggest that local membrane charge modulates the interaction between β<sub>2</sub>AR and competing G protein subtypes.

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