title: Disruption of the lipid-transporting LdMT-LdRos3 complex in Leishmania donovani affects membrane lipid asymmetry but not host cell invasion.
abstract: Maintenance and regulation of the asymmetric lipid distribution across eukaryotic plasma membranes is governed by the concerted action of specific membrane proteins controlling lipid movement across the bilayer. Here, we show that the miltefosine transporter (LdMT), a member of the P4-ATPase subfamily in Leishmania donovani, and the Cdc50-like protein LdRos3 form a stable complex that plays an essential role in maintaining phospholipid asymmetry in the parasite plasma membrane. Loss of either LdMT or LdRos3 abolishes ATP-dependent transport of NBD-labelled phosphatidylethanolamine (PE) and phosphatidylcholine from the outer to the inner plasma membrane leaflet and results in an increased cell surface exposure of endogenous PE. We also find that promastigotes of L. donovani lack any detectable amount of phosphatidylserine (PS) but retain their infectivity in THP-1-derived macrophages. Likewise, infectivity was unchanged for parasites without LdMT-LdRos3 complexes. We conclude that exposure of PS and PE to the exoplasmic leaflet is not crucial for the infectivity of L. donovani promastigotes.
authors.affiliation: Institute of Biology and Biophysics, Humboldt-University Berlin, Berlin, Germany.
authors.forename: Adrien
authors.initials: A
authors.lastname: Weingärtner
authors.forename: Björn
authors.initials: B
authors.lastname: Drobot
authors.forename: Andreas
authors.initials: A
authors.lastname: Herrmann
authors.forename: María P
authors.initials: MP
authors.lastname: Sánchez-Cañete
authors.forename: Francisco
authors.initials: F
authors.lastname: Gamarro
authors.forename: Santiago
authors.initials: S
authors.lastname: Castanys
authors.forename: Thomas
authors.initials: T
authors.lastname: Günther Pomorski
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D001692:Biological Transport; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D006801:Humans; D007893:Leishmania donovani / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D007898:Leishmaniasis, Visceral / Q000469:parasitology*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D011485:Protein Binding; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D014774:Virulence
chemical_list: D026901:Membrane Transport Proteins; D010743:Phospholipids; D015800:Protozoan Proteins; D000251:Adenosine Triphosphatases; C480230:miltefosine transporter, Leishmania donovani