Mitochondria of Malaria Parasites as a Drug Target

Mitochondria are organelle, which is found in most eukaryotic cells, and play an important roll in production of many biosynthetic intermediates as well as energy transduction. Recently, it has been reported that mitochondria contribute to cellular stress responses such as apoptosis and autophagy. These functions of mitochondria are known to be essential for survival and maintenance of homeostasis. The mitochondria of malaria parasites are quite different from those of their vertebrate hosts. Because these differences markedly contribute to drug selectivity, we have focused on the Plasmodium mitochondrion to develop antimalarial drugs. Here we summarize recent advances in our knowledge of the mitochondria of malaria parasites and discuss future prospective antimalarial drugs targeting the parasite mitochondrion

Malaria Parasites Hijack Host Receptors From Exosomes to Capture Lipoproteins

Malaria parasites cannot multiply in host erythrocytes without cholesterol because they lack complete sterol biosynthesis systems. This suggests parasitized red blood cells (pRBCs) need to capture host sterols, but its mechanism remains unknown. Here we identified a novel high-density lipoprotein (HDL)-delivery pathway operating in blood-stage Plasmodium. In parasitized mouse plasma, exosomes positive for scavenger receptor CD36 and platelet-specific CD41 increased. These CDs were detected in pRBCs and internal parasites. A low molecular antagonist for scavenger receptors, BLT-1, blocked HDL uptake to pRBCs and suppressed Plasmodium growth in vitro. Furthermore, platelet-derived exosomes were internalized in pRBCs. Thus, we presume CD36 is delivered to malaria parasites from platelets by exosomes, which enables parasites to steal HDL for cholesterol supply. Cholesterol needs to cross three membranes (RBC, parasitophorous vacuole and parasite’s plasma membranes) to reach parasite, but our findings can explain the first step of sterol uptake by intracellular parasites

Function of Platelet Glycosphingolipid Microdomains/Lipid Rafts

Lipid rafts are dynamic assemblies of glycosphingolipids, sphingomyelin, cholesterol, and specific proteins which are stabilized into platforms involved in the regulation of vital cellular processes. The rafts at the cell surface play important functions in signal transduction. Recent reports have demonstrated that lipid rafts are spatially and compositionally heterogeneous in the single-cell membrane. In this review, we summarize our recent data on living platelets using two specific probes of raft components: lysenin as a probe of sphingomyelin-rich rafts and BCθ as a probe of cholesterol-rich rafts. Sphingomyelin-rich rafts that are spatially and functionally distinct from the cholesterol-rich rafts were found at spreading platelets. Fibrin is translocated to sphingomyelin-rich rafts and platelet sphingomyelin-rich rafts act as platforms where extracellular fibrin and intracellular actomyosin join to promote clot retraction. On the other hand, the collagen receptor glycoprotein VI is known to be translocated to cholesterol-rich rafts during platelet adhesion to collagen. Furthermore, the functional roles of platelet glycosphingolipids and platelet raft-binding proteins including G protein-coupled receptors, stomatin, prohibitin, flotillin, and HflK/C-domain protein family, tetraspanin family, and calcium channels are discussed

Malaria Parasites Hijack Host Receptors From Exosomes to Capture Lipoproteins

Malaria parasites cannot multiply in host erythrocytes without cholesterol because they lack complete sterol biosynthesis systems. This suggests parasitized red blood cells (pRBCs) need to capture host sterols, but its mechanism remains unknown. Here we identified a novel high-density lipoprotein (HDL)-delivery pathway operating in blood-stage Plasmodium. In parasitized mouse plasma, exosomes positive for scavenger receptor CD36 and platelet-specific CD41 increased. These CDs were detected in pRBCs and internal parasites. A low molecular antagonist for scavenger receptors, BLT-1, blocked HDL uptake to pRBCs and suppressed Plasmodium growth in vitro. Furthermore, platelet-derived exosomes were internalized in pRBCs. Thus, we presume CD36 is delivered to malaria parasites from platelets by exosomes, which enables parasites to steal HDL for cholesterol supply. Cholesterol needs to cross three membranes (RBC, parasitophorous vacuole and parasite’s plasma membranes) to reach parasite, but our findings can explain the first step of sterol uptake by intracellular parasites

MOESM1 of Suppression of experimental cerebral malaria by disruption of malate:quinone oxidoreductase

Additional file 1. Fumarate cycle in Plasmodium falciparum. Fumarate, which is generated via purine biosynthesis, is converted into malate by fumarate hydratase (FH) [11]. Then, malate is converted to oxaloacetate (OOA) by malate:quinone oxidoreductase (MQO) [12]; the oxidation of malate to OOA generates ubiquinol (UQH2), which feeds the electron transport chain [11, 12]. Two of the eight mitochondrial TCA cycle enzymes, FH and MQO, may be essential for survival of asexual-blood-stage P. falciparum [9]. Note: MQO is conserved among all apicomplexan parasites, including all Cryptosporidium species [29]

MOESM4 of Suppression of experimental cerebral malaria by disruption of malate:quinone oxidoreductase

Additional file 4. Deficiency of FH and MQO has no effect on gametocyte production. Blood was obtained from infected mice showing 3% parasitaemia and cultured for 22 h under standardized in vitro culture conditions. Then, mature gametocytes and schizonts were collected by Nycodenz density-gradient centrifugation. (A and B) Expression of gametocyte-specific genes. mdv-1/peg3 [21] and g377 [22] were subjected to semi-quantitative RT–PCR using specific primers (see Additional files 2, 3). The hsp70 was used as a positive control. Samples treated with DNase-treated RNA template (hsp70 (-)) were used as a negative control that is the control of eventual DNA contamination of the RNA preparations. Experiments were performed in duplicate and representative data are shown. (C) Control and Δfh parasites-infected erythrocytes cultured for 22 h. (D) Control and Δmqo parasites-infected erythrocytes cultured for 22 h. White arrows indicate representative mature gametocytes. The scale bars indicate 20 μm. Note that sex-specific features such as nuclear enlargement, the distribution of pigment granules throughout the cytoplasm and enlargement of the cells are observed in both Δfh- and Δmqo-parasite cultures just the same as reported by Mons [23]

Novel Characteristics of Mitochondrial Electron Transport Chain from Eimeria tenella

Eimeria tenella is an intracellular apicomplexan parasite, which infects cecal epithelial cells from chickens and causes hemorrhagic diarrhea and eventual death. We have previously reported the comparative RNA sequence analysis of the E. tenella sporozoite stage between virulent and precocious strains and showed that the expression of several genes involved in mitochondrial electron transport chain (ETC), such as type II NADH dehydrogenase (NDH-2), complex II (succinate:quinone oxidoreductase), malate:quinone oxidoreductase (MQO), and glycerol-3-phosphate dehydrogenase (G3PDH), were upregulated in virulent strain. To study E. tenella mitochondrial ETC in detail, we developed a reproducible method for preparation of mitochondria-rich fraction from sporozoites, which maintained high specific activities of dehydrogenases, such as NDH-2 followed by G3PDH, MQO, complex II, and dihydroorotate dehydrogenase (DHODH). Of particular importance, we showed that E. tenella sporozoite mitochondria possess an intrinsic ability to perform fumarate respiration (via complex II) in addition to the classical oxygen respiration (via complexes III and IV). Further analysis by high-resolution clear native electrophoresis, activity staining, and nano-liquid chromatography tandem-mass spectrometry (nano-LC-MS/MS) provided evidence of a mitochondrial complex II-III-IV supercomplex. Our analysis suggests that complex II from E. tenella has biochemical features distinct to known orthologues and is a potential target for the development of new anticoccidian drugs