Extended Data Fig. 2: Effect of 5D3-Fab on ABCG2 function.
From: Cryo-EM structures of a human ABCG2 mutant trapped in ATP-bound and substrate-bound states
a, Analytical SEC profile of the nanodisc-reconstituted ABCG2EQ–E1S complex in the presence of 5 mM ATP and 5 mM MgCl2. ‘1’ denotes the peak collected. Inset: non-reducing SDS–PAGE of the complex, showing bands for ABCG2 (G2), 5D3-Fab (Fab) and nanodisc (ND). b, ATPase activity of liposome-reconstituted ABCG2, in the presence or absence of 5D3-Fab, and with 0–300 µM E1S. The basal ATPase activity has been normalized (norm) to 0. c, As for b, but with the maximal ATPase activity set to 100%. Each point represents the mean rate derived from technical replicates. For G2 n = 6, except in the case of 0 and 200 µM E1S, for which n = 9. For G2 + Fab, n = 3. d, ATPase activities of ABCG2 in the presence and absence of 5D3-Fab, and either 0 or 50 µM E1S. e, As for d, but with activities in the presence of E1S set to 100%. Bars show means and dots show the rates derived from each technical replicate (same batch of liposomes). Error bars show the standard deviation. f, The EC50 of E1S ATPase stimulation determined using the curves in b and c with the error of the fit (standard deviation) shown. PL, proteoliposome.
