RBC membrane biomechanics and Plasmodium falciparum invasion: probing beyond ligand–receptor interactions

2022 ◽  
Author(s):  
Patrice V. Groomes ◽  
Usheer Kanjee ◽  
Manoj T. Duraisingh
Keyword(s):  
Plasmodium Falciparum ◽  
Rbc Membrane ◽  
Receptor Interactions

Ability of Plasmodium falciparum to invade Southeast Asian ovalocytes varies between parasite lines

Blood ◽  
2004 ◽  
Vol 104 (9) ◽  
pp. 2961-2966 ◽  
Author(s):  
Alfred Cortés ◽  
Ariadna Benet ◽  
Brian M. Cooke ◽  
John W. Barnwell ◽  
John C. Reeder
Keyword(s):  
Plasmodium Falciparum ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Clinical Observation ◽  
Southeast Asian ◽  
Parasite Line ◽  
Invasion Pathways ◽  
Receptor Interactions ◽  
Multiple Ligand

Abstract Plasmodium falciparum, the causative agent of the most lethal form of human malaria, uses multiple ligand-receptor interactions to invade host red blood cells (RBCs). We studied the invasion of P falciparum into abnormal RBCs from humans carrying the Southeast Asian ovalocytosis (SAO) trait. One particular parasite line, 3D7-A, invaded these cells efficiently, whereas all other lines studied invaded SAO RBCs to only about 20% of the extent of normal (non-SAO) cells. This result is consistent with the clinical observation that SAO individuals can experience high-density P falciparum infections and provides an explanation for previous discrepant results on invasion of SAO RBCs. Characterization of the invasion phenotype of 3D7-A revealed that efficient invasion of SAO RBCs was paralleled by relatively efficient invasion of normal RBCs treated with either neuraminidase, trypsin, or chymotrypsin and a novel capacity to invade normal RBCs treated sequentially with both neuraminidase and trypsin. Our results suggest that only parasites able to use some particular invasion pathways can invade SAO RBCs efficiently in culture. A similar situation might occur in the field.

scholarly journals A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyte

2012 ◽  
Vol 6 (06) ◽  
pp. 536-541 ◽  
Author(s):  
Emanuela Ferru ◽  
Anntonella Pantaleo ◽  
Francesco Turrini
Keyword(s):  
Plasmodium Falciparum ◽  
Membrane Surface ◽  
Electrophoretic Analysis ◽  
Cytoskeletal Proteins ◽  
Surface Proteins ◽  
New Method ◽  
Major Drawback ◽  
Rbc Membrane ◽  
Wide Range ◽  
Erythrocyte Surface

Introduction: We propose a new method for the selective labeling, isolation and electrophoretic analysis of the Plasmodium falciparum protein exposed on the erythrocyte cell surface. Historically, membrane surface proteins have been isolated using a surface biotinylation followed by capture of biotin-conjugated protein via an avidin/streptavidin-coated solid support. The major drawback of the standard methods has been the labeling of internal proteins due to fast internalization of biotin. Methodology: To solve this problem, we used a biotin label that does not permeate through the membrane. As a further precaution to avoid the purification of non surface exposed proteins, we directly challenged whole labeled cells with avidin coated beads and then solubilized them using non ionic detergents. Results: A marked enrichment of most of the RBC membrane proteins known to face the external surface of the membrane validated the specificity of the method; furthermore, only small amounts of haemoglobin and cytoskeletal proteins were detected. A wide range of P. falciparum proteins were additionally described to be exposed on the erythrocyte surface. Some of them have been previously observed and used as vaccine candidates while a number of newly described antigens have been presently identified. Those antigens require further characterization and validation with additional methods. Conclusion: Surface proteins preparations were very reproducible and identification of proteins by mass spectrometry has been demonstrated to be feasible and effective.

scholarly journals Functional Analysis of the Exported Type IV HSP40 Protein PfGECO in Plasmodium falciparum Gametocytes

2011 ◽  
Vol 10 (11) ◽  
pp. 1492-1503 ◽  
Author(s):  
Belinda J. Morahan ◽  
Carolyn Strobel ◽  
Uzma Hasan ◽  
Beata Czesny ◽  
Pierre-Yves Mantel ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Heat Shock ◽  
Protective Role ◽  
Targeted Disruption ◽  
Content Type ◽  
Type Iv ◽  
Rbc Membrane ◽  
Host Defense Response ◽  
Hsp40 Protein

ABSTRACTDuringPlasmodium falciparuminfection, host red blood cell (RBC) remodeling is required for the parasite's survival. Such modifications are mediated by the export of parasite proteins into the RBC that alter the architecture of the RBC membrane and enable cytoadherence. It is probable that some exported proteins also play a protective role against the host defense response. This may be of particular importance for the gametocyte stage of the life cycle that is responsible for malaria transmission, since the gametocyte remains in contact with blood as it proceeds through five morphological stages (I to V) during its 12-day maturation. Using microarray analysis, we identified several genes with encoded secretory or export sequences that were differentially expressed during early gametocytogenesis. One of these,PfGECO, encodes a predicted type IV heat shock protein 40 (HSP40) that we show is expressed in gametocyte stages I to IV and is exported to the RBC cytoplasm. HSPs are traditionally induced under stressful conditions to maintain homeostasis, butPfGECOexpression was not increased upon heat shock, suggesting an alternate function. Targeted disruption ofPfGECOindicated that the gene is not essential for gametocytogenesisin vitro, and quantitative reverse transcriptase PCR (RT-PCR) showed that there was no compensatory expression of the other type IV HSP40 genes. AlthoughP. falciparumHSP40 members are implicated in the trafficking of proteins to the RBC surface, removal of PfGECO did not affect the targeting of other exported gametocyte proteins. This work has expanded the repertoire of known gametocyte-exported proteins to include a type IV HSP40, PfGECO.

scholarly journals Plasmodium falciparum malaria: association of knobs on the surface of infected erythrocytes with a histidine-rich protein and the erythrocyte skeleton.

1984 ◽  
Vol 98 (4) ◽  
pp. 1256-1264 ◽  
Author(s):  
J H Leech ◽  
J W Barnwell ◽  
M Aikawa ◽  
L H Miller ◽  
R J Howard
Keyword(s):  
Plasmodium Falciparum ◽  
Subcellular Location ◽  
Plasmodium Falciparum Malaria ◽  
Rbc Membrane ◽  
Thin Section Electron Microscopy ◽  
Detergent Extraction ◽  
Section Electron ◽  
Original Observation ◽  
Triton X ◽  
Section Electron Microscopy

Plasmodium falciparum-infected erythrocytes (RBC) develop surface protrusions (knobs) which consist of electron-dense submembrane cups and the overlying RBC plasma membrane. Knobs mediate cytoadherence to endothelial cells. Falciparum variants exist that lack knobs. Using knobby (K+) and knobless (K-) variants of two strains of P. falciparum, we confirmed Kilejian's original observation that a histidine-rich protein occurred in K+ parasites but not K- variants (Kilejian, A., 1979, Proc. Natl. Acad. Sci. USA, 76:4650-4653; and Kilejian, A., 1980, J. Exp. Med., 151:1534-1538). Two additional histidine-rich proteins of lower molecular weight were synthesized by K+ and K- variants of both strains. We used differential detergent extraction and thin-section electron microscopy to investigate the subcellular location of the histidine-rich protein unique to K+ parasites. Triton X-100, Zwittergent 314, cholic acid, CHAPS, and Triton X-100/0.6 M KCl failed to extract the unique histidine-rich protein. The residues insoluble in these detergents contained the unique histidine-rich protein and electron-dense cups. The protein was extracted by 1% SDS and by 1% Triton X-100/9 M urea. The electron-dense cups were missing from the insoluble residues of these detergents. The electron-dense cups and the unique histidine-rich protein appeared to be associated with the RBC skeleton, particularly RBC protein bands 1, 2, 4.1, and 5. We propose that the unique histidine-rich protein binds to the RBC skeleton to form the electron-dense cup. The electron-dense cup produces knobs by forming focal protrusions of the RBC membrane. These protrusions are the specific points of attachment between infected RBC and endothelium.

scholarly journals Novel Antimalarial Aminoquinolines: Heme Binding and Effects on Normal or Plasmodium falciparum-Parasitized Human Erythrocytes

2009 ◽  
Vol 53 (10) ◽  
pp. 4339-4344 ◽  
Author(s):  
Fausta Omodeo-Salè ◽  
Lucia Cortelezzi ◽  
Nicoletta Basilico ◽  
Manolo Casagrande ◽  
Anna Sparatore ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Antimalarial Activity ◽  
Heme Binding ◽  
Human Red Blood Cells ◽  
Rbc Membrane ◽  
Alkyl Derivatives ◽  
Resistant Strains ◽  
Dose Dependent

ABSTRACT Two new quinolizidinyl-alkyl derivatives of 7-chloro-4-aminoquinoline, named AM-1 and AP4b, which are highly effective in vitro against both the D10 (chloroquine [CQ] susceptible) and W2 (CQ resistant) strains of Plasmodium falciparum and in vivo in the rodent malaria model, have been studied for their ability to bind to and be internalized by normal or parasitized human red blood cells (RBC) and for their effects on RBC membrane stability. In addition, an analysis of the heme binding properties of these compounds and of their ability to inhibit beta-hematin formation in vitro has been performed. Binding of AM1 or AP4b to RBC is rapid, dose dependent, and linearly related to RBC density. Their accumulation in parasitized RBC (pRBC) is increased twofold compared to levels in normal RBC. Binding of AM1 or AP4b to both normal and pRBC is higher than that of CQ, in agreement with the lower pKa and higher lipophilicity of the compounds. AM1 or AP4b is not hemolytic per se and is less hemolytic than CQ when hemolysis is accelerated (induced) by hematin. Moreover, AM-1 and AP4b bind heme with a stoichiometry of interaction similar to that of CQ (about 1:1.7) but with a lower affinity. They both inhibit dose dependently the formation of beta-hematin in vitro with a 50% inhibitory concentration comparable to that of CQ. Taken together, these results suggest that the antimalarial activity of AM1 or AP4b is likely due to inhibition of hemozoin formation and that the efficacy of these compounds against the CQ-resistant strains can be ascribed to their hydrophobicity and capacity to accumulate in the vacuolar lipid (elevated lipid accumulation ratios).

scholarly journals Carrier-Mediated Partitioning of Artemisinin into Plasmodium falciparum-Infected Erythrocytes

2002 ◽  
Vol 46 (1) ◽  
pp. 105-109 ◽  
Author(s):  
Nehal Vyas ◽  
Bonnie A. Avery ◽  
Mitchell A. Avery ◽  
Christy M. Wyandt
Keyword(s):  
Plasmodium Falciparum ◽  
Drug Concentration ◽  
Blood Cells ◽  
Competitive Inhibition ◽  
Temperature Dependent ◽  
Optimum Time ◽  
Rbc Membrane ◽  
Drug Concentrations ◽  
Infected Rbcs ◽  
Time Periods

ABSTRACT The purpose of the present study was to characterize the partitioning of artemisinin into both uninfected and Plasmodium falciparum-infected red blood cells (RBCs). The partitioning of [14C](+)-artemisinin into RBCs was studied at four different hematocrit levels and eight time periods. At the optimum time of 2 h, the partitioning process was investigated with eight different drug concentrations ranging from 0.88 to 3.52 μM at 37 and 4°C. The effect of the presence of unlabeled artemisinin on the partitioning of the same concentration of [14C]artemisinin was studied. About 35 to 40% of the drug was seen to partition into uninfected RBCs at a hematocrit of 33%, irrespective of the incubation period or the drug concentration used. In contrast, infected RBCs showed an increase in partitioning of the drug with time until saturation was achieved at 1 h. While the partitioning of artemisinin into parasitized RBCs at 37°C was found to be significantly higher than that in nonparasitized RBCs, at 4°C both parasitized and nonparasitized RBCs showed identical partitioning of the drug. The partitioning of [14C]artemisinin into parasitized RBCs was completely inhibited in the presence of the same concentration of unlabeled artemisinin. However, no such effect was observed in nonparasitized cells, and no evidence suggesting that binding of the drug in parasitized RBCs is reversible was found. The partitioning of artemisinin into parasitized RBCs was found to be rapid, saturable, temperature dependent, irreversible, and subject to competitive inhibition with unlabeled artemisinin. The results obtained suggest the involvement of carrier mediation in the partitioning of artemisinin across the parasitized RBC membrane. In contrast, simple passive diffusion of artemisinin was seen in nonparasitized RBCs.

scholarly journals The hydration state of human red blood cells and their susceptibility to invasion by Plasmodium falciparum

Blood ◽  
2005 ◽  
Vol 105 (12) ◽  
pp. 4853-4860 ◽  
Author(s):  
Teresa Tiffert ◽  
Virgilio L. Lew ◽  
Hagai Ginsburg ◽  
Miriam Krugliak ◽  
Laure Croisille ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Protective Factor ◽  
Gene Frequencies ◽  
Protection Mechanism ◽  
Human Red Blood Cells ◽  
Rbc Membrane ◽  
Healthy Donors ◽  
Hydration State ◽  
High Gene ◽  
Cell Fractions

Abstract In most inherited red blood cell (RBC) disorders with high gene frequencies in malaria-endemic regions, the distribution of RBC hydration states is much wider than normal. The relationship between the hydration state of circulating RBCs and protection against severe falciparum malaria remains unexplored. The present investigation was prompted by a casual observation suggesting that falciparum merozoites were unable to invade isotonically dehydrated normal RBCs. We designed an experimental model to induce uniform and stable isotonic volume changes in RBC populations from healthy donors by increasing or decreasing their KCl contents through a reversible K+ permeabilization pulse. Swollen and mildly dehydrated RBCs were able to sustain Plasmodium falciparum cultures with similar efficiency to untreated RBCs. However, parasite invasion and growth were progressively reduced in dehydrated RBCs. In a parallel study, P falciparum invasion was investigated in density-fractionated RBCs from healthy subjects and from individuals with inherited RBC abnormalities affecting primarily hemoglobin (Hb) or the RBC membrane (thalassemias, hereditary ovalocytosis, xerocytosis, Hb CC, and Hb CS). Invasion was invariably reduced in the dense cell fractions in all conditions. These results suggest that the presence of dense RBCs is a protective factor, additional to any other protection mechanism prevailing in each of the different pathologies. (Blood. 2005; 105:4853-4860)

Characterisation of Plasmodium falciparum RESA-like protein peptides that bind specifically to erythrocytes and inhibit invasion

2007 ◽  
Vol 388 (1) ◽  
pp. 15-24 ◽  
Author(s):  
Luis Eduardo Rodriguez ◽  
Ricardo Vera ◽  
John Valbuena ◽  
Hernando Curtidor ◽  
Javier Garcia ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Membrane Surface ◽  
Binding Assays ◽  
Gene Encoding ◽  
Trophozoite Stage ◽  
Rbc Membrane ◽  
Erythrocyte Invasion ◽  
Erythrocyte Surface ◽  
Maurer's Clefts

Abstract The Plasmodium falciparum ring-erythrocyte surface antigen (RESA)-like putative protein was identified and characterised. PCR and RT-PCR assays revealed that the gene encoding this protein was both present and being transcribed in P. falciparum strain FCB-2 16 h after erythrocyte invasion. Indirect immunofluorescence studies detected this protein in infected erythrocyte (IE) cytosol in dense fluorescent granules similar to Maurer's clefts at 16–20 h (parasites in ring and trophozoite stages) and very strongly on IE membranes at 22 h, suggesting that it is synthesised during early ring stages (16 h) and transported to the infected red blood cell (RBC) membrane surface during the trophozoite stage (22 h). Western blotting showed that antisera produced against polymerised synthetic peptides of this protein recognised a 72-kDa band in P. falciparum schizont lysate. P. falciparum RESA-like peptides used in normal RBC binding assays revealed that peptides 30326 (101NAEKI LGFDD KNILE ALDLFY120), 30334 (281RVTWK KLRTK MIKAL KKSLTY300) and 30342 (431SSPQR LKFTA GGGFC GKLRNY450) bind with high activity and saturability, presenting nM affinity constants. These peptides contain α-helical structural elements, as determined by circular dichroism, and inhibit P. falciparum in vitro invasion of normal RBCs by up to 91%, suggesting that some RESA-like protein regions are involved in intra-erythrocyte stage P. falciparum invasion.

scholarly journals Breakdown in membrane asymmetry regulation leads to monocyte recognition of P. falciparum-infected red blood cells

2021 ◽  
Vol 17 (2) ◽  
pp. e1009259
Author(s):  
Merryn Fraser ◽  
Weidong Jing ◽  
Stefan Bröer ◽  
Florian Kurth ◽  
Leif-Erik Sander ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Fluorescent Dyes ◽  
Host Immune System ◽  
Enzymatic Reactions ◽  
Membrane Asymmetry ◽  
Outer Leaflet ◽  
Rbc Membrane ◽  
Infected Rbcs ◽  
Underlying Causes

The human malaria parasite Plasmodium falciparum relies on lipids to survive; this makes its lipid metabolism an attractive drug target. The lipid phosphatidylserine (PS) is usually confined to the inner leaflet of the red blood cell membrane (RBC) bilayer; however, some studies suggest that infection with the intracellular parasite results in the presence of this lipid in the RBC membrane outer leaflet, where it could act as a recognition signal to phagocytes. Here, we used fluorescent lipid analogues and probes to investigate the enzymatic reactions responsible for maintaining asymmetry between membrane leaflets, and found that in parasitised RBCs the maintenance of membrane asymmetry was partly disrupted, and PS was increased in the outer leaflet. We examined the underlying causes for the differences between uninfected and infected RBCs using fluorescent dyes and probes, and found that calcium levels increased in the infected RBC cytoplasm, whereas membrane cholesterol was depleted from the erythrocyte plasma membrane. We explored the resulting effect of PS exposure on enhanced phagocytosis by monocytes, and show that infected RBCs must expend energy to limit phagocyte recognition, and provide experimental evidence that PS exposure contributes to phagocytic recognition of P. falciparum-infected RBCs. Together, these findings underscore the pivotal role for PS exposure on the surface of Plasmodium falciparum-infected erythrocytes for in vivo interactions with the host immune system, and provide a rationale for targeted antimalarial drug design.

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