Transbilayer Dynamics of Phospholipids in the Plasma Membrane of the Leishmania Genus

Marcos Gonzaga Dos Santos,Sandra Marcia Muxel,Lucile Maria Floeter-Winter,Thomas Günther Pomorski,Ricardo Andrade Zampieri

doi:10.1371/journal.pone.0055604

Marcos Gonzaga Dos Santos, Sandra Marcia Muxel + Show 3 more

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https://doi.org/10.1371/journal.pone.0055604

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Abstract

Protozoans of the Leishmania genus are the etiological agents of a wide spectrum of diseases commonly known as leishmaniases. Lipid organization of the plasma membrane of the parasite may mimic the lipid organization of mammalian apoptotic cells and play a role in phagocytosis and parasite survival in the mammal host. Here, we analyzed the phospholipid dynamics in the plasma membrane of both the L. (Leishmania) and the L. (Viannia) subgenera. We found that the activity and substrate specificity of the inward translocation machinery varied between Leishmania species. The differences in activity of inward phospholipid transport correlated with the different sensitivities of the various species towards the alkyl-phospholipid analogue miltefosine. Furthermore, all species exhibited a phospholipid scramblase activity in their plasma membranes upon stimulation with calcium ionophores. However, binding of annexin V to the parasite surface was only detected for a subpopulation of parasites during the stationary growth phase and only marginally enhanced by scramblase activation.

Highlights

Protozoans of the genus Leishmania cause a complex disease called leishmaniases, whose clinical manifestations have been divided into three principal types that exhibit different degrees of severity and mortality: cutaneous, mucocutaneous, and visceral [1]
Phospholipid transport activity in the plasma membrane of Leishmania parasites To compare the activity and characteristics of lipid transporters in the plasma membrane of various Leishmania species promastigotes, we first examined the amount of inwardly transported 7nitrobenz-2-oxa-1,3-diazole (NBD)-labeled phospholipids, which is protected against albumin extraction after the translocation, by flow cytometry
For L. (L.) amazonensis, L. (L.) chagasi, and L. (L.) donovani, rapid and selective internalization of NBD-PC and NBD-PS were observed with transport rates at least 24- and 12-fold higher, respectively, than NBD-SM (Fig. 1A)

Summary

Introduction

Protozoans of the genus Leishmania cause a complex disease called leishmaniases, whose clinical manifestations have been divided into three principal types that exhibit different degrees of severity and mortality: cutaneous, mucocutaneous, and visceral [1]. Species of this genus alternate between an intracellular amastigote stage in the mammalian host and a free, flagellated promastigote stage in the gut of the sand fly vector. A critical point in the host-parasite interaction involves the attachment of the parasite to the intestinal epithelium and a posterior detachment and migration to the proboscis of the insect vector. Recent results suggest that Leishmania parasites lack PS [6,7,8,9], the detection of cell surface lipids by staining with labeled annexin V, a protein that interacts with PS and with phosphatidic acid and phosphatidylinositol-4,5bisphosphate [10], is consistent with the hypothesis that the protozoan parasite might take advantage of a regulated surfacedisplay of specific lipids to invade and survive in host macrophages

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