ABCMdb

PMID: 17537873

Glavinas H, Kis E, Pal A, Kovacs R, Jani M, Vagi E, Molnar E, Bansaghi S, Kele Z, Janaky T, Bathori G, von Richter O, Koomen GJ, Krajcsi P
ABCG2 (breast cancer resistance protein/mitoxantrone resistance-associated protein) ATPase assay: a useful tool to detect drug-transporter interactions.
Drug Metab Dispos. 2007 Sep;35(9):1533-42. Epub 2007 May 30., [PubMed]

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No. Mutations Sentence Comment

24 ABCG2 p.Arg482Gly 9 Copyright (c) 2007 by The American Society for Pharmacology and Experimental Therapeutics 14605/3238592 DMD 35:1533-1542, 2007 Printed in U.S.A. 1533 The first ABCG2 ATPase assays using Sf9 membranes were reported on the R482G version of the transporter (Ozvegy et al., 2001). Login to comment 45 ABCG2 p.Lys86Met Human membrane vesicle preparations containing ABCG2 (MXR-M) and control human membrane preparations (M-CTRL), as well as insect cell membranes containing the human transporter (MXR-Sf9) and control insect membranes (beta-gal-Sf9-CTRL, MXR-K86M-Sf9-CTRL), were obtained from SOLVO Biotechnology (Budapest, Hungary, http://www. Login to comment 47 ABCG2 p.Lys86Met The insect membrane vesicle preparations were obtained using recombinant baculoviruses encoding wild-type human ABCG2, inactive ABCG2-K86M mutant (carrying a mutation at a crucial position of the catalytic center of ATP binding and cleavage), and beta-galactosidase (Ozvegy et al., 2001, 2002). Login to comment 102 ABCG2 p.Lys86Met ABCG2 p.Lys86Met Sf9 preparations containing the wild-type or the defective (K86M) transporter (MXR-Sf9, lane 1; MXR-K86M-Sf9-CTRL, lane 2) displayed a strong band with apparent molecular mass of 55 to 60 kDa. Login to comment 110 ABCG2 p.Lys86Met The transport of [3 H]methotrexate could not be observed in Sf9 membranes containing the K86M- FIG. 2. Login to comment 111 ABCG2 p.Lys86Met Vanadate-sensitive ATPase activity of MXR-Sf9 (A, C) and MXR-M (B, D) and defMXR-K86M-Sf9 (E) and M-CTRL (F) preparations in the presence of ABCG2 substrates and inhibitors at different concentrations. Membranes containing 20 ␮g of total protein were incubated at 37°C for 40 min in the presence of different concentrations of test compounds. Login to comment 125 ABCG2 p.Lys86Met Sf9 membranes expressing the defective (MXR-K86M-Sf9-CTRL) version of ABCG2 show similar baseline vanadate-sensitive ATPase activity, which was not modulated by any of the substrates tested (Fig. 2E). Login to comment 181 ABCG2 p.Lys86Met The vanadate-sensitive ATPase activity of MXR-Sf9 membranes in the presence of Ko143 or Ko134 and the vanadate-sensitive ATPase activity present of membranes containing the defective transporter (MXR-K86M-Sf9-CTRL) show that the Sf9 preparations contain some (ϳ10 nmol Pi/mg/min) non-ABCG2-related vanadate-sensitive FIG. 5. Login to comment