MAEBL Plasmodium falciparum protein peptides bind specifically to erythrocytes and inhibit in vitro merozoite invasion
Section snippets
Materials and methods
Peptide synthesis. One hundred and three sequential 20-mer peptides, corresponding to the complete P. falciparum MAEBL protein amino acid sequence [GenBank Accession No. NP_701342], were synthesised by the solid phase multiple peptide system [16], [17]; t-Boc amino acids (Bachem); and MBHA resin (0.7 meq/g) were used. Peptides were cleaved by the low-high HF technique [18], purified by RP-HPLC, lyophilised, and analysed by MALDI-TOF mass spectrometry. Tyrosine was added to those peptides which
MAEBL peptides bind specifically to human erythrocytes
Binding assays were used to determine specific erythrocyte binding activity for 103 synthetic peptides covering the total length of the MAEBL protein [GenBank Accession No. NP_701342]. Peptide binding activity was defined as being the amount (pmol) of peptide that bound specifically to erythrocyte per added peptide (pmol). High activity binding peptides (HABPs) were defined as those peptides showing activity greater than or equal to 2%, according to previously established criteria [20], [21],
Discussion
A better understanding of the complex process of P. falciparum merozoite invasion requires that the numerous potential parasite ligands become identified and characterised. Our studies have required an innovative methodology to be employed in the search for sequences which can be precisely modified for use as candidates for a vaccine against malaria [28], [29], [30], [31]. Some of the important ligand determinants for invasion belong to the members of the ebl family [11]. Differences in
Acknowledgements
This research project was supported by the President of the Republic of Colombia’s office and the Colombian Ministry of Public Health. We thank Jason Garry for reading the manuscript.
References (40)
- et al.
The Duffy receptor family of Plasmodium knowlesi is located within the micronemes of invasive malaria merozoites
Cell
(1990) - et al.
The apical organelles of malaria merozoites: host cell selection, invasion, host immunity and immune evasion
Microbes Infect.
(2000) - et al.
Apical organelles of Apicomplexa: biology and isolation by subcellular fractionation
Mol. Biochem. Parasitol.
(2001) - et al.
Erythrocyte binding protein homologues of rodent malaria parasites
Mol. Biochem. Parasitol.
(1997) - et al.
Plasmodium falciparum MAEBL is a unique member of the ebl family
Mol. Biochem. Parasitol.
(2002) - et al.
An expanding ebl family of Plasmodium falciparum
Trends Parasitol.
(2001) - et al.
Spatial and temporal dynamics of the secretory pathway during differentiation of the Plasmodium yoelii schizont
Mol. Biochem. Parasitol.
(2000) - et al.
Quantitative analysis of drug-receptor interactions. 1. Determination of kinetic and equilibrium properties
Life Sci.
(1981) - et al.
A flow cytometric method for assessing viability of intraerythrocytic hemoparasites
J. Immunol. Methods
(1991) - et al.
Identification, expression, and functional characterization of MAEBL, a sporozoite and asexual blood stage chimeric erythrocyte binding protein of Plasmodium falciparum
Mol. Biochem. Parasitol.
(2002)
Glycophorin B as an EBA-175 independent Plasmodium falciparum receptor on human erythrocytes
Mol. Biochem. Parasitol.
Invasion of erythrocytes by Plasmodium falciparum malaria parasites: evidence for receptor heterogeneity and two receptors
Blood
Erythrocyte receptor recognition varies in Plasmodium falciparum isolates
Mol. Biochem. Parasitol.
Molecular insights into receptors used by malaria parasites for erythrocyte invasion
Curr. Opin. Hematol.
Invasion of erythrocytes by malaria merozoites
Science
Erythrocyte entry by malarial parasites. A moving junction between erythrocyte and parasite
J. Cell Biol.
The invasion of erythrocytes by malarial merozoites
Lamellar membranes associated with rhoptries in erythrocytic merozoites of Plasmodium knowlesi: a clue to the mechanism of invasion
Parasitology
A family of erythrocyte binding proteins of malaria parasites
Proc. Natl. Acad. Sci. USA
A family of chimeric erythrocyte binding proteins of malaria parasites
Proc. Natl. Acad. Sci. USA
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2017, Biochemical and Biophysical Research CommunicationsFunctional, structural, and immunological compartmentalisation of malaria invasive proteins
2007, Biochemical and Biophysical Research CommunicationsCitation Excerpt :Only the MSP-1 19 kDa fragment anchored to the merozoite via a GPI tail (where HABP 5501 was localised) is found inside newly invaded RBC [33]. Regarding MAEBL (presenting a hybrid structure between AMA protein and EBL proteins) [34], the first two conserved HABPs (30181 and 30195) localised in the region having the greatest homology with AMA have β-turn, strand or unordered configurations, as happens with conserved AMA protein HABPs. The remaining conserved HABPs had predominantly α-helical structures, as happens with EBA and EBL proteins (Table 1).
Synthetic peptides from Plasmodium falciparum apical membrane antigen 1 (AMA-1) specifically interacting with human hepatocytes
2006, BiochimieCitation Excerpt :These results suggest that the inhibiting effect could be due to peptides interacting with similar proteins on the erythrocyte membrane, agreeing with the homology presented by sequences amongst themselves. In fact, it has been reported that peptides 30209 and 30181 bind to a 32 kDa protein [38], this being a similar weight to that found for peptide 4316. Cross-linking studies for HABP 4321 showed that this peptide bound two proteins whose molecular weights were 36 and 58 kDa, suggesting a possible receptor for AMA-1 protein in the HepG2 cell line.
Cellular adhesive phenomena in apicomplexan parasites of red blood cells
2005, Veterinary ParasitologyIdentifying Plasmodium falciparum merozoite surface protein-10 human erythrocyte specific binding regions
2005, BiochimieCitation Excerpt :Merozoite binding to erythrocytes requires parasite receptors [3]. Receptor-ligand interactions using synthetic peptides have revealed that many proteins from Plasmodium falciparum sexual and hepatic stages appear to be involved in the complex invasion process of human hepatocytes [4–7], reticulocytes [8–10] and erythrocytes [11–20]. Although very little is known about the particular role of any individual protein, it has been postulated that merozoite surface proteins (MSP-1 [21], MSP-2 [22], MSP-4 [23], MSP-5 [24] and MSP-8 [17]) are involved in initial merozoite interaction with the erythrocyte surface [22–27].