The Yeast Plasma Membrane ATP Binding Cassette (ABC) Transporter Aus1
| Autor: | Peter Müller, David L. Daleke, Magdalena Marek, Gunnar Dittmar, Gabriele Schreiber, Andreas Herrmann, Sigrid Milles, Thomas Güenther Pomorski |
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| Rok vydání: | 2011 |
| Předmět: |
biology
Saccharomyces cerevisiae Walker motifs Transporter ATP-binding cassette transporter Cell Biology biology.organism_classification Biochemistry Sterol Cell biology Cell membrane chemistry.chemical_compound medicine.anatomical_structure chemistry ATP hydrolysis medicine Molecular Biology Adenosine triphosphate |
| Zdroj: | Journal of Biological Chemistry. 286:21835-21843 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m111.244525 |
| Popis: | The ATP binding cassette (ABC) transporter Aus1 is expressed under anaerobic growth conditions at the plasma membrane of the yeast Saccharomyces cerevisiae and is required for sterol uptake. These observations suggest that Aus1 promotes the translocation of sterols across membranes, but the precise transport mechanism has yet to be identified. In this study, an extraction and purification procedure was developed to characterize the Aus1 transporter. The detergent-solubilized protein was able to bind and hydrolyze ATP. Mutagenesis of the conserved lysine to methionine in the Walker A motif abolished ATP hydrolysis. Likewise, ATP hydrolysis was inhibited by classical inhibitors of ABC transporters. Upon reconstitution into proteoliposomes, the ATPase activity of Aus1 was specifically stimulated by phosphatidylserine (PS) in a stereoselective manner. We also found that Aus1-dependent sterol uptake, but not Aus1 expression and trafficking to the plasma membrane, was affected by changes in cellular PS levels. These results suggest a direct interaction between Aus1 and PS that is critical for the activity of the transporter. |
| Databáze: | OpenAIRE |
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