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Dualsteric GPCR targeting: a novel route to binding and signaling pathway selectivity
Author(s) -
Antony Johannes,
Kellershohn Kerstin,
MohrAndrä Marion,
Kebig Anna,
Prilla Stefanie,
Muth Mathias,
Heller Eberhard,
Disingrini Teresa,
Dallanoce Clelia,
Bertoni Simona,
Schrobang Jasmin,
Trankle Christian,
Kostenis Evi,
Christopoulos Arthur,
Höltje HansDieter,
Barocelli Elisabetta,
Amici Marco,
Holzgrabe Ulrike,
Mohr Klaus
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/fj.08-114751
Subject(s) - allosteric regulation , g protein coupled receptor , chemistry , functional selectivity , receptor , muscarinic acetylcholine receptor m5 , muscarinic acetylcholine receptor m2 , microbiology and biotechnology , signal transduction , muscarinic acetylcholine receptor , muscarinic acetylcholine receptor m3 , biochemistry , biology
Selective modulation of cell function by G protein‐coupled receptor (GPCR) activation is highly desirable for basic research and therapy but difficult to achieve. We present a novel strategy toward this goal using muscarinic acetylcholine receptors as a model. The five subtypes bind their physiological transmitter in the highly conserved orthosteric site within the transmembrane domains of the receptors. Orthosteric muscarinic activators have no binding selectivity and poor signaling specificity. There is a less well conserved allosteric site at the extracellular entrance of the binding pocket. To gain subtype‐selective receptor activation, we synthesized two hybrids fusing a highly potent oxotremorine‐like orthosteric activator with M 2 ‐selective bis(ammonio)alkane‐type allosteric fragments. Radioligand binding in wild‐type and mutant receptors supplemented by receptor docking simulations proved M 2 selective and true allosteric/orthosteric binding. G protein activation measurements using orthosteric and allosteric blockers identified the orthosteric part of the hybrid to engender receptor activation. Hybrid‐induced dynamic mass redistribution in CHO‐hM 2 cells disclosed pathway‐specific signaling. Selective receptor activation (M 2 >M 1 >M 3 ) was verified in living tissue preparations. As allosteric sites are increasingly recognized on GPCRs, the dualsteric concept of GPCR targeting represents a new avenue toward potent agonists for selective receptor and signaling pathway activation.— Antony, J., Kellershohn, K., Mohr‐Andrä, M., Kebig, A., Prilla, S., Muth, M., Heller, E., Disingrini, T., Dallanoce, C., Bertoni, S., Schrobang, J., Tränkle, C., Kostenis, E., Christopoulos, A., Höltje, H.‐D., Barocelli, E., De Amici, M., Holzgrabe, U., Mohr, K. Dualsteric GPCR targeting: a novel route to binding and signaling pathway selectivity. FASEB J. 23, 442–450 (2009)

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