Potent and Selective Peptide-based Inhibition of the G Protein Gαq

Charpentier, Thomas H.; Waldo, Gary L.; Lowery-Gionta, Emily G.; Krajewski, Krzysztof; Strahl, Brian D.; Kash, Thomas L.; Harden, T. Kendall; Sondek, John

doi:10.1074/jbc.m116.740407

Cited by 26 publications

(42 citation statements)

References 40 publications

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“…4D), indicating that PLCβ3 is required for LPA chemotaxis. To verify this result, we also used a selective peptide inhibitor of the Gαq subunit (Charpentier et al, 2016). The peptide, a GFP tagged fragment of PLCβ3 harboring an I860A mutation, has a high affinity for Gαq and functions as a dominant negative for Gαq signaling, including the activation of PLCbs.…”

Section: Plcβ3 and Pkcα Are Required For Lpa Chemotaxismentioning

confidence: 98%

Lysophosphatidic acid provokes fibroblast chemotaxis through combinatorial regulation of myosin II

Asokan

Johnson

Sondek

et al. 2018

Preprint

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Lysophophatidic acid (LPA), a biologically active phospholipid that is ubiquitously present in tissues and organs, provokes cellular responses such as proliferation, apoptosis, differentiation and migration via activation of G-protein coupled receptors. These receptors activate a broad range of intracellular signaling cascades to mediate these responses. Using microfluidic chambers that generate and maintain stable gradients, we observed that chemotaxis of fibroblasts to LPA has higher directional fidelity than chemotaxis provoked by the receptor tyrosine kinase (RTK) ligand platelet-derived growth factor (PDGF). Unlike fast moving amoeboid cells, mesenchymal cells such as fibroblasts do not require PI3K for chemotaxis to a GPCR ligand. In addition, the Arp2/3 complex is not required for fibroblast GPCR-based chemotaxis in either 2D or 3D environments. Our data indicate that combinatorial regulation of myosin II involving global activation by RhoA/ROCK and local inhibition of myosin II at the leading edge by PKC results in highly efficient chemotaxis of fibroblasts to LPA. Based on these observations, we develop a simple mathematical model to explain how dual regulation of myosin II is responsible for enhanced chemotaxis in LPA gradients relative to PDGF. Using pharmacological approaches, we test predictions of this model and modulate the fidelity of LPA and PDGF chemotaxis.

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Section: Plcβ3 and Pkcα Are Required For Lpa Chemotaxismentioning

confidence: 98%

Lysophosphatidic acid provokes fibroblast chemotaxis through combinatorial regulation of myosin II

Asokan

Johnson

Sondek

et al. 2018

Preprint

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“…With a potent Gαs inhibitor, GsIN-1, functioning in vitro, we next asked whether GsIN-1 could penetrate cell membranes, as peptide-based chemical probes often suffer from poor cell 8 permeability. Several G protein-specific linear peptides exhibit in vitro activities but have no reported cellular efficacy, likely due to their low cell permeability (22)(23)(24)(25)(26)(27). In order to quantitatively evaluate the cell permeability of GsIN-1, we used a recently developed HaloTagbased assay known as a chloroalkane penetration assay (CAPA) (Fig.…”

Section: The Cyclic Peptide Gsin-1 Is Able To Penetrate Hela Cell Memmentioning

confidence: 99%

State-selective Modulation of Heterotrimeric Gαs Signaling with Macrocyclic Peptides

Dai

Gao

et al. 2020

Preprint

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The G protein-coupled receptor (GPCR) cascade leading to production of the second messenger cAMP is replete with pharmacologically targetable receptors and enzymes with the exception of the stimulatory G protein α subunit, Gαs. GTPases remain largely undruggable given the difficulty of displacing high affinity guanine nucleotides and the lack of other drug binding sites. We 5 explored a chemical library of 10 12 cyclic peptides in order to expand the chemical search for inhibitors of this enzyme class. We identified a macrocyclic peptide, GsIN-1, that antagonizes the GTP-bound active state of Gαs with high G protein specificity and nucleotide-binding-state selectivity. A 1.57 Å co-crystal structure reveals that GsIN-1 binds to a novel switch II / α3 pocket in Gαs and directly blocks adenylyl cyclase activation. GsIN-1 inhibits isoproterenol-stimulated 10 Gαs activation through binding to the crystallographically defined pocket. The discovery of GsIN-1 provides path for the development of state-dependent GTPase inhibitors. 15 20 One sentence summary: Targeting the previously undruggable ON-state of the GTPase Gαs with a macrocyclic peptide inhibitor.

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“…The structure was confirmed by mass spectrometry. A, Modified from Charpentier et al [Color figure can be viewed at wileyonlinelibrary.com]…”

Section: Selective Gq Inhibitorsmentioning

confidence: 99%

Recent achievements in developing selective G_q inhibitors

Zhang

Nielsen

Strømgaard

2019

Medicinal Research Reviews

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G proteins are key mediators of G protein‐coupled receptor (GPCR) signaling, facilitating a plethora of important physiological processes. The role of G proteins is much less understood than other aspects of GPCR function, which is largely due to the shortage of potent and selective G protein inhibitors. The natural cyclic depsipeptides YM‐254890 and FR900359 are two of the very few known selective inhibitors of the Gq subfamily, and are used as unique pharmacological tools in the study of G q‐mediated signaling. Moreover, a peptide‐based G protein antagonist‐2A (GP‐2A), a 27‐residue peptide (27mer(I860A)) derived from phospholipase C‐β3 (PLC‐β3), and the small molecule BIM‐46187 have also been characterized as selective G q inhibitors within the past 5 years. In this review, we highlight the recent development in chemical syntheses, characterization, and mechanism of action of these selective G q inhibitors. The development and application of G q‐selective inhibitors will expand our knowledge of the structure and function of G protein‐mediated signaling, shed light on the development of inhibitors for other G protein classes, and feed in to drug discovery for diseases where G proteins are implicated, including various forms of cancer.

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Potent and Selective Peptide-based Inhibition of the G Protein Gαq

Cited by 26 publications

References 40 publications

Lysophosphatidic acid provokes fibroblast chemotaxis through combinatorial regulation of myosin II

Lysophosphatidic acid provokes fibroblast chemotaxis through combinatorial regulation of myosin II

State-selective Modulation of Heterotrimeric Gαs Signaling with Macrocyclic Peptides

Recent achievements in developing selective G_q inhibitors

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Potent and Selective Peptide-based Inhibition of the G Protein Gαq

Cited by 26 publications

References 40 publications

Lysophosphatidic acid provokes fibroblast chemotaxis through combinatorial regulation of myosin II

Lysophosphatidic acid provokes fibroblast chemotaxis through combinatorial regulation of myosin II

State-selective Modulation of Heterotrimeric Gαs Signaling with Macrocyclic Peptides

Recent achievements in developing selective Gq inhibitors

Contact Info

Product

Resources

About

Recent achievements in developing selective G_q inhibitors