DAPLE protein inhibits nucleotide exchange on Gαs and Gαq via the same motif that activates Gαi

DAPLE protein inhibits nucleotide exchange on Gαs and Gαq via the same motif that activates GαiJournal of Biological ChemistryVol. 295Issue 8PreviewBesides being regulated by G-protein–coupled receptors, the activity of heterotrimeric G proteins is modulated by many cytoplasmic proteins. GIV/Girdin and DAPLE (Dvl-associating protein with a high frequency of leucine) are the best-characterized members of a group of cytoplasmic regulators that contain a Gα-binding and -activating (GBA) motif and whose dysregulation underlies human diseases, including cancer and birth defects. GBA motif–containing proteins were originally reported to modulate G proteins by binding Gα subunits of the Gi/o family (Gαi) over other families (such as Gs, Gq/11, or G12/13), and promoting nucleotide exchange in vitro. Full-Text PDF Open Access VOLUME 295 (2020) PAGES 2270-2284 MgCl2 was omitted as a reagent in two buffers described under “In vitro protein-binding assays with GST-fused proteins” section (under “Experimental procedures”). “Beads were washed twice with PBS and resuspended in 300 µl of binding buffer (50 mm Tris-HCl (pH 7.4), 100 mm NaCl, 0.4% (vol:vol) NP-40, 5 mm EDTA, 2 mm DTT)” should read as “Beads were washed twice with PBS and resuspended in 300 µl of binding buffer (50 mm Tris-HCl (pH 7.4), 100 mm NaCl, 10 mm MgCl2, 0.4% (vol:vol) NP-40, 5 mm EDTA, 2 mm DTT).” “Beads were washed four times with 1 ml of wash buffer (4.3 mm Na2HPO4, 1.4 mm KH2PO4, pH 7.4, 137 mm NaCl, 2.7 mm KCl, 0.1% (vol:vol) Tween-20, 5 mm EDTA, 1 mm DTT)” should read as “Beads were washed four times with 1 ml of wash buffer (4.3 mm Na2HPO4, 1.4 mm KH2PO4, pH 7.4, 137 mm NaCl, 2.7 mm KCl, 0.1% (vol:vol) Tween-20, 10 mm MgCl2, 5 mm EDTA, 1 mm DTT).”