GONUTS has been updated to MW1.31 Most things seem to be working but be sure to report problems.

Have any questions? Please email us at ecoliwiki@gmail.com

PMID:17098188

From GONUTS
Jump to: navigation, search
Citation

McDevitt, CA, Collins, RF, Conway, M, Modok, S, Storm, J, Kerr, ID, Ford, RC and Callaghan, R (2006) Purification and 3D structural analysis of oligomeric human multidrug transporter ABCG2. Structure 14:1623-32

Abstract

ABCG2 is a multidrug efflux pump associated with resistance of cancer cells to a plethora of unrelated drugs. ABCG2 is a "half-transporter," and previous studies have indicated that it forms homodimers and higher oligomeric species. In this manuscript, electron microscopic structural analysis directly addressed this issue. An N-terminal hexahistidine-tagged ABCG2(R482G) isoform was expressed to high levels in insect cells. An extensive detergent screen was employed to effect extraction of ABCG2(R482G) from membranes and identified only the fos-choline detergents as efficient. Soluble protein was purified to >95% homogeneity by a three-step procedure while retaining the ability to bind substrates. Cryonegative stain electron microscopy of purified ABCG2(R482G) provided 3D structural data at a resolution of approximately 18 A. Single-particle analysis revealed that the complex forms a tetrameric complex ( approximately 180 A in diameter x approximately 140 A high) with an aqueous central region. We interpret the tetrameric structure as comprising four homodimeric ABCG2(R482G) complexes.

Links

PubMed Online version:10.1016/j.str.2006.08.014

Keywords

ATP Binding Cassette Transporter, Subfamily G, Member 2; ATP-Binding Cassette Transporters/chemistry; ATP-Binding Cassette Transporters/isolation & purification; Animals; Cell Membrane/metabolism; Chromatography/methods; Cryoelectron Microscopy; Detergents/chemistry; Detergents/pharmacology; Dimerization; Humans; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Insecta; Neoplasm Proteins/chemistry; Neoplasm Proteins/isolation & purification; Protein Conformation; Protein Isoforms; Protein Structure, Tertiary

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:ABCG2

enables

GO:0042802: identical protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UNQ0

F

Seeded From UniProt

complete

Notes

See also

References

See Help:References for how to manage references in GONUTS.