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Proc Natl Acad Sci U S A. 2014 Jul 22;111(29):10744-8. doi: 10.1073/pnas.1410415111. Epub 2014 Jul 8.

Covalent agonists for studying G protein-coupled receptor activation.

Author information

1
Department of Chemistry and Pharmacy, Friedrich Alexander University, 91052 Erlangen, Germany; and.
2
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305.
3
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305 kobilka@stanford.edu peter.gmeiner@fau.de.
4
Department of Chemistry and Pharmacy, Friedrich Alexander University, 91052 Erlangen, Germany; and kobilka@stanford.edu peter.gmeiner@fau.de.

Abstract

Structural studies on G protein-coupled receptors (GPCRs) provide important insights into the architecture and function of these important drug targets. However, the crystallization of GPCRs in active states is particularly challenging, requiring the formation of stable and conformationally homogeneous ligand-receptor complexes. Native hormones, neurotransmitters, and synthetic agonists that bind with low affinity are ineffective at stabilizing an active state for crystallogenesis. To promote structural studies on the pharmacologically highly relevant class of aminergic GPCRs, we here present the development of covalently binding molecular tools activating Gs-, Gi-, and Gq-coupled receptors. The covalent agonists are derived from the monoamine neurotransmitters noradrenaline, dopamine, serotonin, and histamine, and they were accessed using a general and versatile synthetic strategy. We demonstrate that the tool compounds presented herein display an efficient covalent binding mode and that the respective covalent ligand-receptor complexes activate G proteins comparable to the natural neurotransmitters. A crystal structure of the β2-adrenoreceptor in complex with a covalent noradrenaline analog and a conformationally selective antibody (nanobody) verified that these agonists can be used to facilitate crystallogenesis.

KEYWORDS:

chemical biology; chemical probes; structural biology

PMID:
25006259
PMCID:
PMC4115510
DOI:
10.1073/pnas.1410415111
[Indexed for MEDLINE]
Free PMC Article

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