P-type atpase apt1 of the fungal pathogen Cryptococcus neoformans is a lipid flippase of broad substrate specificity
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P-type atpase apt1 of the fungal pathogen Cryptococcus neoformans is a lipid flippase of broad substrate specificity. / Stanchev, Lyubomir Dimitrov; Rizzo, Juliana; Peschel, Rebecca; Pazurek, Lilli A.; Bredegaard, Lasse; Veit, Sarina; Laerbusch, Sabine; Rodrigues, Marcio L.; López-Marqués, Rosa L.; Pomorski, Thomas Günther.
In: Journal of Fungi, Vol. 7, No. 10, 843, 2021.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - P-type atpase apt1 of the fungal pathogen Cryptococcus neoformans is a lipid flippase of broad substrate specificity
AU - Stanchev, Lyubomir Dimitrov
AU - Rizzo, Juliana
AU - Peschel, Rebecca
AU - Pazurek, Lilli A.
AU - Bredegaard, Lasse
AU - Veit, Sarina
AU - Laerbusch, Sabine
AU - Rodrigues, Marcio L.
AU - López-Marqués, Rosa L.
AU - Pomorski, Thomas Günther
N1 - Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021
Y1 - 2021
N2 - 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.
AB - 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.
KW - CDC50 protein
KW - Heterologous expression
KW - Lipid transport
KW - Membrane transport protein
KW - P4-ATPase
KW - β-subunit
U2 - 10.3390/jof7100843
DO - 10.3390/jof7100843
M3 - Journal article
C2 - 34682264
AN - SCOPUS:85117326816
VL - 7
JO - Journal of Fungi
JF - Journal of Fungi
SN - 2309-608X
IS - 10
M1 - 843
ER -
ID: 284197742