Extended Data Fig. 6: Contributions of residues at the Best2-GS interacting interface.
From: Bestrophin-2 and glutamine synthetase form a complex for glutamate release
(a) Bar chart showing GS activities in the whole-cell lysates of HEK293 cells transiently transfected with indicated hBest2 and hGS constructs; n = 3 for each group. (b) FRET two-hybrid analysis of the interaction between Cerulean-tagged hGS and Venus-tagged hBest2 constructs. Bar chart showing 1/Kd,EFF of the interaction; n = 3 for each group; *p = 0.01, 0.01, 0.02 and 0.01 for the WT-mutant pairs from left to right, respectively, compared to the WT-WT pair. (c) Bar chart showing GS activities in the cytosolic lysates of HEK293 cells transiently transfected with indicated hGS constructs; n = 3 for each group; *p = 9E-3 and 0.02 for untransfected and hGS R299E transfected cells, respectively, compared to cells transfected with WT hGS. (d) Population steady-state current density-voltage relationships of hBest2 in the presence of hGS G23A or K52A mutant (magenta triangles, n = 7 for each group), compared to hBest2 without (black triangle, n = 9) or with WT hGS (red circle, n = 5) at 1 μM [Ca2+]i in transiently transfected HEK293 cells; *p = 0.04 compared to currents from cells expressing hBest2 alone. All error bars are presented as mean values +/− SEM; p values are calculated by two-tailed unpaired Student’s t test.
