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The membrane‐proximal C‐terminal Arg/Lys cluster mediates G protein‐coupled receptor interaction with tubulin and is required for receptor transport to the cell surface
Author(s) -
Duvernay Matthew Thomas,
Zhang Xiaoping,
Dong Chunmin,
Wu Guangyu
Publication year - 2009
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.23.1_supplement.945.5
Subject(s) - tubulin , microtubule , microbiology and biotechnology , receptor , g protein coupled receptor , chemistry , docking (animal) , endoplasmic reticulum , nocodazole , biochemistry , biology , cell , cytoskeleton , medicine , nursing
A growing body of evidence supports a pivotal role for microtubules in intracellular protein trafficking. However, the role of microtubule network in biosynthesis of G protein‐coupled receptor (GPCR) remains largely unknown. By using peptide‐conjugated Agarose affinity column and proteomics, we identified α and β‐tubulin interacting with the α 2B ‐adrenergic receptor (AR) C‐terminus (CT), which was confirmed by GST fusion protein pull down assay using purified tubulin or brain cytosolic extract. Site‐directed mutagenesis of the CT revealed the interaction was mediated by a cluster of basic residues in helix 8, suggestive of an ionic interaction, consistent with the negatively charged EExEEY motif in tubulin as a common docking site for microtubule‐associated proteins. Indeed, NaCl dose‐dependently inhibited α 2B ‐AR interaction with tubulin. Interestingly, the CT of α 2B ‐AR and angiotensin II type I receptor (AT1R), but not β 2 ‐AR, physically associated with tubulin and mutation of the Arg/Lys cluster in α 2B ‐AR and AT1R abolished their export and ERK1/2 activation and the mutated receptors were extensively arrested in the endoplasmic reticulum (ER). Finally, nocodazole treatment blocked α 2B ‐AR cell surface expression by 60%. These data indicate for the first time that the Arg/Lys cluster‐directed GPCR interaction with tubulin coordinates receptor export from the ER to the plasma membrane (GM076167).