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https://www.arca.fiocruz.br/handle/icict/50002
P-TYPE ATPASE APT1 OF THE FUNGAL PATHOGEN CRYPTOCOCCUS NEOFORMANS IS A LIPID FLIPPASE OF BROAD SUBSTRATE SPECIFICITY
Lipídeos
Proteínas de Membrana Transportadoras
ATPases do Tipo-P
Virulência
Author
Affilliation
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany. / Department of Plant and Environmental Sciences. University of Copenhagen. Frederiksberg, Denmark.
Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Paulo de Góes. Rio de Janeiro, RJ, Brasil. / Institut Pasteur. Département de Mycologie. Unité Biologie des ARN des Pathogènes Fongiques. Paris, France.
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany.
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany.
Department of Plant and Environmental Sciences. University of Copenhagen. Frederiksberg, Denmark.
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany.
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany.
Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Paulo de Góes. Rio de Janeiro, RJ, Brasil. / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Department of Plant and Environmental Sciences. University of Copenhagen. Frederiksberg, Denmark.
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany. / Department of Plant and Environmental Sciences. University of Copenhagen. Frederiksberg, Denmark.
Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Paulo de Góes. Rio de Janeiro, RJ, Brasil. / Institut Pasteur. Département de Mycologie. Unité Biologie des ARN des Pathogènes Fongiques. Paris, France.
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany.
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany.
Department of Plant and Environmental Sciences. University of Copenhagen. Frederiksberg, Denmark.
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany.
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany.
Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Paulo de Góes. Rio de Janeiro, RJ, Brasil. / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Department of Plant and Environmental Sciences. University of Copenhagen. Frederiksberg, Denmark.
Department of Molecular Biochemistry. Faculty of Chemistry and Biochemistry. Ruhr University Bochum. Bochum, Germany. / Department of Plant and Environmental Sciences. University of Copenhagen. Frederiksberg, Denmark.
Abstract
Lipid flippases of the P4-ATPase family are ATP-driven transporters that translocate lipids from the exoplasmic to the cytosolic leaflet of biological membranes. In the encapsulated fungal pathogen Cryptococcus neoformans, the P4-ATPase Apt1p is an important regulator of polysaccharide secretion and pathogenesis, but its biochemical characterization is lacking. Phylogenetic analysis revealed that Apt1p belongs to the subclade of P4A-ATPases characterized by the common requirement for a β-subunit. Using heterologous expression in S. cerevisiae, we demonstrate that Apt1p forms a heterodimeric complex with the C. neoformans Cdc50 protein. This association is required for both localization and activity of the transporter complex. Lipid flippase activity of the heterodimeric complex was assessed by complementation tests and uptake assays employing fluorescent lipids and revealed a broad substrate specificity, including several phospholipids, the alkylphospholipid miltefosine, and the glycolipids glucosyl- and galactosylceramide. Our results suggest that transbilayer lipid transport in C. neoformans is finely regulated to promote fungal virulence, which reinforces the
potential of Apt1p as a target for antifungal drug development.
DeCS
Cryptococcus neoformansLipídeos
Proteínas de Membrana Transportadoras
ATPases do Tipo-P
Virulência
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