The human breast cancer resistance protein (BCRP/ABCG2) shows conformational changes with mitoxantrone.
BCRP/ABCG2 mediates efflux of drugs and xenobiotics. BCRP was expressed in Pichia pastoris, purified to > 90% homogeneity, and subjected to two-dimensional (2D) crystallization. The 2D crystals showed a p12(1) symmetry and projection maps were determined at 5 A resolution by cryo-electron microscopy. Two crystal forms with and without mitoxantrone were observed with unit cell dimensions of a = 55.4 A, b = 81.4 A, gamma = 89.8 degrees , and a = 57.3 A, b = 88.0 A, gamma = 89.7 degrees , respectively. The projection map without mitoxantrone revealed an asymmetric structure with ring-shaped density features probably corresponding to a bundle of transmembrane alpha helices, and appeared more open and less symmetric than the map with mitroxantrone. The open and closed inward-facing forms of BCRP were generated by homology modeling, representing the substrate-free and substrate-bound conformations in the absence of nucleotide, respectively. These models are consistent with the experimentally observed conformational change upon substrate binding.
Rosenberg
Mark F
MF
Manchester Interdisciplinary Biocentre, 131 Princess Street, University of Manchester, Manchester, M1 7DN, UK. mark.rosenberg@manchester.ac.uk
Bikadi
Zsolt
Z
Chan
Janice
J
Liu
Xiaoping
X
Ni
Zhanglin
Z
Cai
Xiaokun
X
Ford
Robert C
RC
Mao
Qingcheng
Q
D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000235:genetics*; D000970:Antineoplastic Agents / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D020285:Cryoelectron Microscopy / Q000379:methods; D018360:Crystallography, X-Ray; D000939:Epitopes / Q000737:chemistry; D006801:Humans; D008854:Microscopy, Electron / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D008942:Mitoxantrone / Q000737:chemistry / Q000494:pharmacology*; D008958:Models, Molecular; D008968:Molecular Conformation; D009363:Neoplasm Proteins / Q000737:chemistry* / Q000235:genetics*; D010843:Pichia; D011487:Protein Conformation; D017433:Protein Structure, Secondary
C116614:ABCG2 protein, human; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D000970:Antineoplastic Agents; D000939:Epitopes; D009363:Neoplasm Proteins; D008942:Mitoxantrone
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