Functional Significance of Conserved Cysteines in the Extracellular Loops of the ATP Binding Cassette Transporter Pdr11p
| Autor: | Mara Klöhn, Feng Xian, Gunnar Dittmar, Daniele Silvestro, Rosa L. López-Marqués, Thomas Günther Pomorski, Lyubomir Dimitrov Stanchev, Magdalena Marek |
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| Rok vydání: | 2020 |
| Předmět: |
Microbiology (medical)
Protein trafficking Mutant Saccharomyces cerevisiae Cell ATP-binding cassette transporter disulfide bonds Plant Science sterol uptake ATPase activity Article Sterol uptake 03 medical and health sciences ABC transport proteins medicine Extracellular Disulfide bonds lcsh:QH301-705.5 Ecology Evolution Behavior and Systematics 030304 developmental biology 0303 health sciences biology Chemistry 030302 biochemistry & molecular biology Transporter biology.organism_classification Yeast medicine.anatomical_structure Biochemistry lcsh:Biology (General) protein trafficking Cysteine |
| Zdroj: | Journal of Fungi Journal of Fungi, Vol 7, Iss 2, p 2 (2021) Stanchev, L D, Marek, M, Xian, F, Klöhn, M, Silvestro, D, Dittmar, G, López-Marqués, R L & Günther Pomorski, T 2021, ' Functional Significance of Conserved Cysteines in the Extracellular Loops of the ATP Binding Cassette Transporter Pdr11p ', Journal of Fungi, vol. 7, no. 1, 2 . https://doi.org/10.3390/jof7010002 Volume 7 Issue 1 |
| ISSN: | 2309-608X |
| DOI: | 10.3390/jof7010002 |
| Popis: | The pleiotropic drug resistance (PDR) transporter Pdr11p is expressed under anaerobic growth conditions at the plasma membrane of the yeast Saccharomyces cerevisiae, where it facilitates the uptake of exogenous sterols. Members of the fungal PDR family contain six conserved cysteines in their extracellular loops (ECL). For the functional analysis of these cysteine residues in Pdr11p, we generated a series of single cysteine-to-serine mutants. All mutant proteins expressed well and displayed robust ATPase activity upon purification. Mass-spectrometry analysis identified two cysteine residues (C582 and C603) in ECL3 forming a disulfide bond. Further characterization by cell-based assays showed that all mutants are compromised in facilitating sterol uptake, protein stability, and trafficking to the plasma membrane. Our data highlight the fundamental importance of all six extracellular cysteine residues for the functional integrity of Pdr11p and provide new structural insights into the PDR family of transporters. |
| Databáze: | OpenAIRE |
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