MESA is a Plasmodium falciparum phosphoprotein associated with the erythrocyte membrane skeleton

doi:10.1016/0166-6851(88)90152-1

Molecular and Biochemical Parasitology

Volume 31, Issue 3, December 1988, Pages 223-231

https://doi.org/10.1016/0166-6851(88)90152-1 Get rights and content

Abstract

The mature-parasite-infected erythrocyte surface antigen (MESA) of Plasmodium falciparum is an antigenically variable, high molecular weight protein of trophozoites and schizonts that is located at the erythrocyte surface membrane. It is first synthesized at the late ring stage and continues to be synthesized until late schizogony. MESA cannot be detected on the external surface of erythrocytes infected by trophozoites and early schizonts but is located at the internal surface in association with the erythrocyte membrane skeleton. The degree of association with the membrane skeleton varies among parasite lines, being greater in knobby parasite lines. MESA is phosphorylated and is present in a similar location to another phosphoprotein, the ring-infected erythtrocyte surface antigen (RESA). However, it differs from RESA in being detected at a later stage of asexual development of the parasite.

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Cited by (64)

The Plasmodium falciparum MESA erythrocyte cytoskeleton-binding (MEC)motif binds to erythrocyte ankyrin
2019, Molecular and Biochemical Parasitology
Citation Excerpt :
Binding of the MESA MEC motif to ANK1 had no effect on erythrocyte rigidity in the microsphere filtration assay (Fig. 9). This is consistent with data from MESA gene deletion strains, which did not reveal any obvious phenotypes [27,28,31]. In addition, the MEC motif binding site, ZU5C, has no known function.
The MESA erythrocyte cytoskeleton binding (MEC) motif is a 13-amino acid sequence found in 14 exported Plasmodium falciparum proteins. First identified in the P. falciparum Mature-parasite-infected Erythrocyte Surface Antigen (MESA), the MEC motif is sufficient to target proteins to the infected red blood cell cytoskeleton. To identify host cell targets, purified MESA MEC motif was incubated with a soluble extract from uninfected erythrocytes, precipitated and subjected to mass spectrometry. The most abundant co-purifying protein was erythrocyte ankyrin (ANK1). A direct interaction between the MEC motif and ANK1 was independently verified using co-purification experiments, the split-luciferase assay, and the yeast two-hybrid assay. A systematic mutational analysis of the core MEC motif demonstrated a critical role for the conserved aspartic acid residue at the C-terminus of the MEC motif for binding to both erythrocyte inside-out vesicles and to ANK1. Using a panel of ANK1 constructs, the MEC motif binding site was localized to the ZU5^C domain, which has no known function. The MEC motif had no impact on erythrocyte deformability when introduced into uninfected erythrocyte ghosts, suggesting the MEC motif’s primary function is to target exported proteins to the cytoskeleton. Finally, we show that PF3D7_0402100 (PFD0095c) binds to ANK1 and band 4.1, likely through its MEC and PHIST motifs, respectively. In conclusion, we have provided multiple lines of evidence that the MEC motif binds to erythrocyte ANK1.
Compositional and expression analyses of the glideosome during the Plasmodium life cycle reveal an additional myosin light chain required for maximum motility
2017, Journal of Biological Chemistry
Myosin A (MyoA) is a Class XIV myosin implicated in gliding motility and host cell and tissue invasion by malaria parasites. MyoA is part of a membrane-associated protein complex called the glideosome, which is essential for parasite motility and includes the MyoA light chain myosin tail domain–interacting protein (MTIP) and several glideosome-associated proteins (GAPs). However, most studies of MyoA have focused on single stages of the parasite life cycle. We examined MyoA expression throughout the Plasmodium berghei life cycle in both mammalian and insect hosts. In extracellular ookinetes, sporozoites, and merozoites, MyoA was located at the parasite periphery. In the sexual stages, zygote formation and initial ookinete differentiation precede MyoA synthesis and deposition, which occurred only in the developing protuberance. In developing intracellular asexual blood stages, MyoA was synthesized in mature schizonts and was located at the periphery of segmenting merozoites, where it remained throughout maturation, merozoite egress, and host cell invasion. Besides the known GAPs in the malaria parasite, the complex included GAP40, an additional myosin light chain designated essential light chain (ELC), and several other candidate components. This ELC bound the MyoA neck region adjacent to the MTIP-binding site, and both myosin light chains co-located to the glideosome. Co-expression of MyoA with its two light chains revealed that the presence of both light chains enhances MyoA-dependent actin motility. In conclusion, we have established a system to study the interplay and function of the three glideosome components, enabling the assessment of inhibitors that target this motor complex to block host cell invasion.
Erythrocyte remodeling by Plasmodium falciparum gametocytes in the human host interplay
2015, Trends in Parasitology
Citation Excerpt :
In gametocyte development, although protein export is active from the onset of differentiation [15,41,42], a direct role in erythrocyte host remodeling for most of the observed proteins awaits experimental confirmation. It is possible, for instance, to speculate that the mature parasite-infected erythrocyte surface antigen (MESA), which interacts with the host cell cytoskeleton in asexual parasites [58–61], has a similar function in sexual stages. Also, the megadalton protein Pf11-1, localized to the host cell membrane during gametocytogenesis [62] and recently included among the PEXEL-negative exported proteins (PNEPs) [63], or the PEXEL/HT-containing protein MAL7P1.171, whose ablation resulted in failure of stage I gametocytes to mature [64], may interact with host membrane components.
The spread of malaria critically relies on the presence of Plasmodium transmission stages – the gametocytes – circulating in the blood of an infected individual, which are taken up by Anopheles mosquitoes. A striking feature of Plasmodium falciparum gametocytes is their long development inside the erythrocytes while sequestered in the internal organs of the human host. Recent studies of the molecular and cellular remodeling of the host erythrocyte induced by P. falciparum during gametocyte maturation are shedding light on how these may affect the establishment and maintenance of sequestration of the immature transmission stages and the subsequent release and circulation of mature gametocytes in the peripheral bloodstream.
An exported kinase (FIKK4.2) that mediates virulence-associated changes in Plasmodium falciparum-infected red blood cells
2014, International Journal for Parasitology
Citation Excerpt :
The anti-FIKK4.2 antibody showed no reactivity with normal, uninfected RBCs or RBCs infected with any of the four transgenic KO parasite clones, strongly suggesting that the fluorescence observed in parental 3D7 parasites was highly specific for FIKK4.2. To rule out the possibility that trafficking or localisation of other exported parasite proteins is affected by deletion of FIKK4.2, we compared RBCs infected with parental 3D7 or FIKK4.2 KO parasites that had been labelled with antibodies raised against KAHRP, MESA and Pf332 as three examples of other very well described P. falciparum-encoded exported proteins (Coppel et al., 1988; Crabb et al., 1997; Glenister et al., 2009). By comparing immunofluorescence images of iRBCs infected with either 3D7 or FIKK KO parasites, we observed no noticeable differences in the expression or localisation of any of RBC membrane skeleton-associated proteins in RBCs infected with the FIKK4.2 KO parasites, suggesting that trafficking and transport of other exported proteins is not grossly affected by the absence of FIKK4.2 (Fig. 3D).
Alteration of the adhesive and mechanical properties of red blood cells caused by infection with the malaria parasite Plasmodium falciparum underpin both its survival and extreme pathogenicity. A unique family of parasite putative exported kinases, collectively called FIKK (Phenylalanine (F) – Isoleucine (I) – Lysine (K) – Lysine (K)), has recently been implicated in these pathophysiological processes, however, their precise function in P. falciparum-infected red blood cells or their likely role in malaria pathogenesis remain unknown. Here, for the first time, we demonstrate that one member of the FIKK family, FIKK4.2, can function as an active kinase and is localised in a novel and distinct compartment of the parasite-infected red blood cell which we have called K-dots. Notably, targeted disruption of the gene encoding FIKK4.2 (fikk4.2) dramatically alters the parasite’s ability to modify and remodel the red blood cells in which it multiplies. Specifically, red blood cells infected with fikk4.2 knockout parasites were significantly less rigid and less adhesive when compared with red blood cells infected with normal parasites from which the transgenic clones had been derived, despite expressing similar levels of the major cytoadhesion ligand, PfEMP1, on the red blood cell surface. Notably, these changes were accompanied by dramatically altered knob-structures on infected red blood cells that play a key role in cytoadhesion which is responsible for much of the pathogenesis associated with falciparum malaria. Taken together, our data identifies FIKK4.2 as an important kinase in the pathogenesis of P. falciparum malaria and strengthens the attractiveness of FIKK kinases as targets for the development of novel next-generation anti-malaria drugs.
The ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum stabilizes spectrin tetramers and suppresses further invasion
2007, Blood
Citation Excerpt :
Following invasion, it is released into the host cell cytosol, where it is phosphorylated18 and becomes associated with the membrane of the newly invaded cell. RESA remains detectable in the infected erythrocytes until about 18 to 24 hours after invasion, when it gradually disappears as MESA appears.19 Spectrin exists in the cell predominantly as an α2β2 tetramer, which has the form of a long, flexible rod, with a contour length of 200 nm.
The malaria parasite Plasmodium falciparum releases the ring-infected erythrocyte surface antigen (RESA) inside the red cell on entry. The protein migrates to the host cell membrane, where it binds to spectrin, but neither the nature of the interaction nor its functional consequences have previously been defined. Here, we identify the binding motifs involved in the interaction and describe a possible function. We have found that spectrin binds to a 108–amino acid fragment (residues 663-770) of RESA, and that this RESA fragment binds to repeat 16 of the β-chain, close to the labile dimer-dimer self-association site. We further show that the RESA fragment stabilizes the spectrin tetramer against dissociation into its constituent dimers, both in situ and in solution. This is accompanied by enhanced resistance of the cell to both mechanical and thermal degradation. Resealed erythrocytes containing RESA_663-770 display resistance to invasion by merozoites of P falciparum. We infer that the evolutionary advantage of RESA to the parasite lies in its ability to prevent invasion of cells that are already host to a developing parasite, as well as possibly to guard the cell against thermal damage at the elevated body temperatures prevailing in febrile crises.
The Hsp40 proteins of Plasmodium falciparum and other apicomplexa: Regulating chaperone power in the parasite and the host
2007, International Journal of Biochemistry and Cell Biology
Citation Excerpt :
Furthermore, data from the disruption of the gene encoding RESA has suggested that RESA plays a role in resistance against heat shock (Silva et al., 2005). Similarly to RESA, the MESA protein (also referred to as P. falciparum erythrocyte membrane protein 2 [PfEMP-2]) associates with the erythrocyte membrane skeleton (Coppel et al., 1988) where it is involved in the formation of the knob structures implicated in the cytoadherence of infected erythrocytes (Sharma, 1991). Three of the hypothetical type IV PEXEL/HT-containing proteins bear potential transmembrane domains (Table 1), and therefore may similarly be implicated in interactions with the erythrocyte membrane.
Extensive structural and functional remodelling of Plasmodium falciparum (malaria)-infected erythrocytes follows the export of a range of proteins of parasite origin (exportome) across the parasitophorous vacuole into the host erythrocyte. The genome of P. falciparum encodes a diverse chaperone complement including at least 43 members of the heat shock protein 40 kDa (Hsp40) family, and six members of the heat shock protein 70 kDa (Hsp70) family. Nearly half of the Hsp40 proteins of P. falciparum are predicted to contain a PEXEL/HT (Plasmodium export element/host targeting signal) sequence motif, and hence are likely to be part of the exportome. In this review we critically evaluate the classification, sequence similarity and clustering, and possible interactors of the P. falciparum Hsp40 chaperone machinery. In addition to the types I, II and III Hsp40 proteins all exhibiting the signature J-domain, the P. falciparum genome also encodes a number of specialized Hsp40 proteins with a J-like domain, which we have categorized as type IV Hsp40 proteins. Analysis of the potential P. falciparum Hsp40 protein interaction network revealed connections predominantly with cytoskeletal and membrane proteins, transcriptional machinery, DNA repair and replication machinery, translational machinery, the proteasome and proteolytic enzymes, and enzymes involved in cellular physiology. Comparison of the Hsp40 proteins of P. falciparum to those of other apicomplexa reveals that most of the proteins (especially the PEXEL/HT-containing proteins) are unique to P. falciparum. Furthermore, very few of the P. falciparum Hsp40 proteins have human homologs, except for those proteins implicated in fundamental biological processes. Our analysis suggests that P. falciparum has evolved an expanded and specialized Hsp40 protein machinery to enable it successfully to invade and remodel the human erythrocyte, and we propose a model in which these proteins are involved in chaperone-mediated translocation, folding, assembly and regulation of parasite and host proteins.

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Molecular and Biochemical Parasitology

Abstract

References (27)

Variable antigen associated with the surface of erythrocytes infected with mature stages of Plasmodium falciparum

Mol. Biochem. Parasitol.

Chromosome size polymorphisms in Plasmodium falciparum can involve deletions and are frequent in natural parasite population

Cell

A tandemly repeated sequence determines the binding domain for an erythrocyte receptor binding protein of P. falciparum

Cell

Cytosolic protein kinase activity associated with the maturation of the malaria parasite Plasmodium berghei

Mol. Biochem. Parasitol.

Plasmodium berghei, P. chabaudi and P. falciparum: similarities in phosphoproteins and protein kinase activities and their stage specific expression

Exp. Parasitol.

Transport of an M_r approx. 300 000 Plasmodium falciparum protein (PfEMP 2) from the intraerythrocytic asexual parasite to the cytoplasmic face of the host cell membrane

J. Cell. Biol.

Two approximately 300 kilodalton Plasmodium falciparum proteins at the surface membrane of infected erythrocytes

Mol. Biochem. Parasitol.

Immune sera recognize on erythrocytes a Plasmodium falciparum antigen composed of repeated amino acid sequences

Nature

Antibodies in malarial sera to parasite antigens in the membrane of erythrocytes infected with early asexual stages of Plasmodium falciparum

J. Exp. Med.

Plasmodium falciparum: Identification and localization of a knob protein antigen expressed by a cDNA clone

Exp. Parasitol.

Human malaria parasites in continuous culture

Science

Synchronization of Plasmodium falciparum erythrocytic stages in culture

J. Parasitol.

Identification of isolate-specific proteins of sorbitol-enriched Plasmodium falciparum infected erythrocytes from Ghambian patients

Parasitology

Cited by (64)

The Plasmodium falciparum MESA erythrocyte cytoskeleton-binding (MEC)motif binds to erythrocyte ankyrin

Compositional and expression analyses of the glideosome during the Plasmodium life cycle reveal an additional myosin light chain required for maximum motility

Erythrocyte remodeling by Plasmodium falciparum gametocytes in the human host interplay

An exported kinase (FIKK4.2) that mediates virulence-associated changes in Plasmodium falciparum-infected red blood cells

The ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum stabilizes spectrin tetramers and suppresses further invasion

The Hsp40 proteins of Plasmodium falciparum and other apicomplexa: Regulating chaperone power in the parasite and the host

MESA is a Plasmodium falciparum phosphoprotein associated with the erythrocyte membrane skeleton

Abstract

Mol. Biochem. Parasitol.

Cell

Cell

Mol. Biochem. Parasitol.

Exp. Parasitol.

Transport of an Mr approx. 300 000 Plasmodium falciparum protein (PfEMP 2) from the intraerythrocytic asexual parasite to the cytoplasmic face of the host cell membrane

J. Cell. Biol.

Two approximately 300 kilodalton Plasmodium falciparum proteins at the surface membrane of infected erythrocytes

Mol. Biochem. Parasitol.

Immune sera recognize on erythrocytes a Plasmodium falciparum antigen composed of repeated amino acid sequences

Nature

Antibodies in malarial sera to parasite antigens in the membrane of erythrocytes infected with early asexual stages of Plasmodium falciparum

J. Exp. Med.

Plasmodium falciparum: Identification and localization of a knob protein antigen expressed by a cDNA clone

Exp. Parasitol.

Human malaria parasites in continuous culture

Science

Synchronization of Plasmodium falciparum erythrocytic stages in culture

J. Parasitol.

Identification of isolate-specific proteins of sorbitol-enriched Plasmodium falciparum infected erythrocytes from Ghambian patients

Parasitology

Transport of an M_r approx. 300 000 Plasmodium falciparum protein (PfEMP 2) from the intraerythrocytic asexual parasite to the cytoplasmic face of the host cell membrane