Fig. 1: Cryo-EM structure of nucleotide-free rMrp2.
From: Structural basis for the modulation of MRP2 activity by phosphorylation and drugs
a Chromatogram and SDS-PAGE analysis of Size Exclusion Chromatography of purified rMrp2. Representatives from three purifications. b Effect of drugs on the basal ATPase activity of rMrp2 reconstituted in destabilised liposomes. The basal ATPase activity of rMrp2 can be stimulated by 1 mM probenecid (PRB) and 0.1 mM methotrexate (MTX). The E1458Q mutant displays no ATPase activity. Results are represented as means with individual data points indicated by asterisc, from three independent experiments. Significantly different as estimated by one way ANOVA (p = 2.32 ×10−7) followed by Tukey test p < 0.001 (red triangle: p < 0.01)). c MRP2 topology with the key domains highlighted. d Ab initio cryo-EM map of rMrp2 at 3.21 Å resolution. Continuous density can be observed for the different rMrp2 domains. The CHS molecules and GDN micelle are shown in pink and transparent grey, respectively. e The nucleotide-free rMrp2 atomic model displays an inward-facing conformation with the TMD cavity occupied by the R-___domain; the helical R-___domain is shown in orange. Source data are provided as a Source Data file.
