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Functional Analysis of Detergent-Solubilized and Membrane-Reconstituted ATP-Binding Cassette Transporters

Poolman, Bert and Doeven, Mark K. and Geertsma, Eric R. and Biemans-Oldehinkel, Esther and Konings, Wil N. and Rees, Douglas C. (2005) Functional Analysis of Detergent-Solubilized and Membrane-Reconstituted ATP-Binding Cassette Transporters. In: Phase II Conjugation Enzymes and Transport Systems. Methods in Enzymology. No.400. Academic Press , San Diego, CA, pp. 429-459. ISBN 978-0-12-182805-9.

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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).

Item Type:Book Section
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Poolman, Bert0000-0002-1455-531X
Rees, Douglas C.0000-0003-4073-1185
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Additional Information:Copyright 2005, Elsevier Inc. All rights reserved. This work was supported by "Top-subsidie" Grant 700-50-302 from the Netherlands Organization for Scientific Research-Chemische Wetenschappen, a Fulbright grant from NACEE, and EU-FP6 (E-Mep programme). We thank Nushin Aghajari, Arnold Driessen, Marc LeMaire, and Gea Schuurman-Wolters for sharing information prior to publication.
Funding AgencyGrant Number
Netherlands Organization for Scientific Research-Chemische Wetenschappen700-50-302
Series Name:Methods in Enzymology
Issue or Number:400
Record Number:CaltechAUTHORS:20141212-144312417
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:52780
Deposited By: SWORD User
Deposited On:22 Dec 2014 21:14
Last Modified:10 Nov 2021 19:43

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