Extended Data Fig. 1: Purification, activity and cryo-EM micrographs of ABCG2.
From: Cryo-EM structures of a human ABCG2 mutant trapped in ATP-bound and substrate-bound states
a, Preparative SEC profile (milli absorbance units (mAU) at 280 nm plotted against retention volume (ml)) of the nanodisc-reconstituted ABCG2EQ–E1S complex. The fraction used for cryo-EM grid preparation is indicated by a red asterisk. Inset: reducing (lane 1) and non-reducing (lane 2) SDS–PAGE of the complex, showing bands for ABCG2 (G2), 5D3-Fab (Fab) and nanodisc (ND). b, Preparative SEC profile of the nanodisc-reconstituted ABCG2EQ–ATP complex. The fraction used for cryo-EM grid preparation is indicated by a red asterisk. Inset: reducing (lane 1) and non-reducing (lane 2) SDS–PAGE of the complex, showing bands for ABCG2 (G2) and nanodisc (ND). c, An example micrograph (drift-corrected, dose-weighted and low-pass-filtered to 20 Å) of the nanodisc-reconstituted ABCG2EQ–E1S sample. White scale bar, 50 nm. d, An example micrograph (drift-corrected, dose-weighted and low-pass-filtered to 20 Å) of the nanodisc-reconstituted ABCG2EQ–ATP sample. White scale bar, 50 nm. e, ATPase activities of nanodisc-reconstituted and E1S-transport activities of liposome-reconstituted ABCG2. In both cases, data for wild-type and mutant (E211Q) ABCG2 are shown. The standard deviation from n technical replicates (same batch of nanodiscs or liposomes) is shown.
