์ฝ˜ํ…์ธ ๋กœ ๊ฑด๋„ˆ๋›ฐ๊ธฐ
Merck

Targeting nucleotide exchange to inhibit constitutively active G protein ฮฑ subunits in cancer cells.

Science signaling (2018-09-06)
Michael D Onken, Carol M Makepeace, Kevin M Kaltenbronn, Stanley M Kanai, Tyson D Todd, Shiqi Wang, Thomas J Broekelmann, Prabakar Kumar Rao, John A Cooper, Kendall J Blumer
์ดˆ๋ก

Constitutively active G protein ฮฑ subunits cause cancer, cholera, Sturge-Weber syndrome, and other disorders. Therapeutic intervention by targeted inhibition of constitutively active Gฮฑ subunits in these disorders has yet to be achieved. We found that constitutively active Gฮฑq in uveal melanoma (UM) cells was inhibited by the cyclic depsipeptide FR900359 (FR). FR allosterically inhibited guanosine diphosphate-for-guanosine triphosphate (GDP/GTP) exchange to trap constitutively active Gฮฑq in inactive, GDP-bound Gฮฑฮฒฮณ heterotrimers. Allosteric inhibition of other Gฮฑ subunits was achieved by the introduction of an FR-binding site. In UM cells driven by constitutively active Gฮฑq, FR inhibited second messenger signaling, arrested cell proliferation, reinstated melanocytic differentiation, and stimulated apoptosis. In contrast, FR had no effect on BRAF-driven UM cells. FR promoted UM cell differentiation by reactivating polycomb repressive complex 2 (PRC2)-mediated gene silencing, a heretofore unrecognized effector system of constitutively active Gฮฑq in UM. Constitutively active Gฮฑq and PRC2 therefore provide therapeutic targets for UM. The development of FR analogs specific for other Gฮฑ subunit subtypes may provide novel therapeutic approaches for diseases driven by constitutively active Gฮฑ subunits or multiple G protein-coupled receptors (GPCRs) where targeting a single receptor is ineffective.

MATERIALS
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Sigma-Aldrich
Anti-Actin Antibody, clone C4, ascites fluid, clone C4, Chemiconยฎ
Sigma-Aldrich
Monoclonal ANTI-FLAGยฎ M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
Sigma-Aldrich
Anti-Histone H3 Antibody, CT, pan, clone A3S, rabbit monoclonal, clone A3S, Upstateยฎ, from rabbit
Sigma-Aldrich
GSK503, ≥98% (HPLC)