title: Functional analysis of detergent-solubilized and membrane-reconstituted ATP-binding cassette transporters.
abstract: ATP-binding cassette (ABC) transporters are vital to any living system and are involved in the translocation of a wide variety of substances, from ions and nutrients to high molecular weight proteins. This chapter describes methods used to purify and membrane reconstitute ABC transporters in a fully functional state. The procedures are largely based on our experience with substrate-binding protein-dependent ABC uptake systems from bacteria, but the approaches should be applicable to multisubunit membrane complexes in general. Also, we present simple methods, based on substrate binding or translocation, to follow the activity of the protein complexes in detergent-solubilized and/or membrane-reconstituted state(s).
authors.affiliation: Department of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute and Materials Science Centre(plus), University of Groningen, The Netherlands.
authors.forename: Bert
authors.initials: B
authors.lastname: Poolman
authors.forename: Mark K
authors.initials: MK
authors.lastname: Doeven
authors.forename: Eric R
authors.initials: ER
authors.lastname: Geertsma
authors.forename: Esther
authors.initials: E
authors.lastname: Biemans-Oldehinkel
authors.forename: Wil N
authors.initials: WN
authors.lastname: Konings
authors.forename: Douglas C
authors.initials: DC
authors.lastname: Rees
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D021961:Cell Membrane Structures / Q000737:chemistry*; D002845:Chromatography; D022162:Cytoplasmic Vesicles / Q000737:chemistry; D003902:Detergents / Q000737:chemistry*; D005453:Fluorescence; D008958:Models, Molecular; D012995:Solubility; D013057:Spectrum Analysis
chemical_list: D018528:ATP-Binding Cassette Transporters; D003902:Detergents