The Gαi-GIV binding interface is a druggable protein-protein interaction

Heterotrimeric G proteins are usually activated by the guanine-nucleotide exchange factor (GEF) activity of GPCRs. However, some non-receptor proteins are also GEFs. GIV (a.k.a Girdin) was the first non-receptor protein for which the GEF activity was ascribed to a well-defined protein sequence that...

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Bibliographic Details
Published in:Scientific reports 2017-08, Vol.7 (1), p.8575-17, Article 8575
Main Authors: DiGiacomo, Vincent, de Opakua, Alain Ibáñez, Papakonstantinou, Maria P., Nguyen, Lien T., Merino, Nekane, Blanco-Canosa, Juan B., Blanco, Francisco J., Garcia-Marcos, Mikel
Format: Article
Language:English
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Amino acid sequence
Binding sites
Cancer
Guanine
High-throughput screening
Humanities and Social Sciences
Magnetic resonance spectroscopy
Metastases
multidisciplinary
NMR
Nuclear magnetic resonance
Protein interaction
Proteins
Science
Science (multidisciplinary)
Spectroscopy
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Summary:Heterotrimeric G proteins are usually activated by the guanine-nucleotide exchange factor (GEF) activity of GPCRs. However, some non-receptor proteins are also GEFs. GIV (a.k.a Girdin) was the first non-receptor protein for which the GEF activity was ascribed to a well-defined protein sequence that directly binds Gαi. GIV expression promotes metastasis and disruption of its binding to Gαi blunts the pro-metastatic behavior of cancer cells. Although this suggests that inhibition of the Gαi-GIV interaction is a promising therapeutic strategy, protein-protein interactions (PPIs) are considered poorly “druggable” targets requiring case-by-case validation. Here, we set out to investigate whether Gαi-GIV is a druggable PPI. We tested a collection of >1,000 compounds on the Gαi-GIV PPI by in silico ligand screening and separately by a chemical high-throughput screening (HTS) assay. Two hits, ATA and NF023, obtained in both screens were confirmed in secondary HTS and low-throughput assays. The binding site of NF023, identified by NMR spectroscopy and biochemical assays, overlaps with the Gαi-GIV interface. Importantly, NF023 did not disrupt Gαi-Gβγ binding, indicating its specificity toward Gαi-GIV. This work establishes the Gαi-GIV PPI as a druggable target and sets the conceptual and technical framework for the discovery of novel inhibitors of this PPI.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-017-08829-7