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Malaria Journal
Published in: Malaria Journal 1/2017

Open Access 01-12-2017 | Research

A novel Pfs38 protein complex on the surface of Plasmodium falciparum blood-stage merozoites

Authors: Gourab Paul, Arunaditya Deshmukh, Inderjeet Kaur, Sumit Rathore, Surbhi Dabral, Ashutosh Panda, Susheel Kumar Singh, Asif Mohmmed, Michael Theisen, Pawan Malhotra

Published in: Malaria Journal | Issue 1/2017

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Abstract

Background

The Plasmodium genome encodes for a number of 6-Cys proteins that contain a module of six cysteine residues forming three intramolecular disulphide bonds. These proteins have been well characterized at transmission as well as hepatic stages of the parasite life cycle. In the present study, a large complex of 6-Cys proteins: Pfs41, Pfs38 and Pfs12 and three other merozoite surface proteins: Glutamate-rich protein (GLURP), SERA5 and MSP-1 were identified on the Plasmodium falciparum merozoite surface.

Methods

Recombinant 6-cys proteins i.e. Pfs38, Pfs12, Pfs41 as well as PfMSP-165 were expressed and purified using Escherichia coli expression system and antibodies were raised against each of these proteins. These antibodies were used to immunoprecipitate the native proteins and their associated partners from parasite lysate. ELISA, Far western, surface plasmon resonance and glycerol density gradient fractionation were carried out to confirm the respective interactions. Furthermore, erythrocyte binding assay with 6-cys proteins were undertaken to find out their possible role in host-parasite infection and seropositivity was assessed using Indian and Liberian sera.

Results

Immunoprecipitation of parasite-derived polypeptides, followed by LC–MS/MS analysis, identified a large Pfs38 complex comprising of 6-cys proteins: Pfs41, Pfs38, Pfs12 and other merozoite surface proteins: GLURP, SERA5 and MSP-1. The existence of such a complex was further corroborated by several protein–protein interaction tools, co-localization and co-sedimentation analysis. Pfs38 protein of Pfs38 complex binds to host red blood cells (RBCs) directly via glycophorin A as a receptor. Seroprevalence analysis showed that of the six antigens, prevalence varied from 40 to 99%, being generally highest for MSP-165 and GLURP proteins.

Conclusions

Together the data show the presence of a large Pfs38 protein-associated complex on the parasite surface which is involved in RBC binding. These results highlight the complex molecular interactions among the P. falciparum merozoite surface proteins and advocate the development of a multi-sub-unit malaria vaccine based on some of these protein complexes on merozoite surface.
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Literature
1.
2.
3.
Ogutu BR, Apollo OJ, McKinney D, Okoth W, Siangla J, Dubovsky F, et al. Blood stage malaria vaccine eliciting high antigen-specific antibody concentrations confers no protection to young children in Western Kenya. PLoS ONE. 2009;4:e4708.CrossRefPubMedPubMedCentral
4.
Sagara I, Dicko A, Ellis RD, Fay MP, Diawara SI, Assadou MH, et al. A randomized controlled phase 2 trial of the blood stage AMA1-C1/Alhydrogel malaria vaccine in children in Mali. Vaccine. 2009;27:3090–8.CrossRefPubMedPubMedCentral
5.
Genton B, Betuela I, Felger I, Al-Yaman F, Anders RF, Saul A, et al. A recombinant blood-stage malaria vaccine reduces Plasmodium falciparum density and exerts selective pressure on parasite populations in a phase 1-2b trial in Papua New Guinea. J Infect Dis. 2002;185:820–7.CrossRefPubMed
6.
Thera MA, Doumbo OK, Coulibaly D, Laurens MB, Ouattara A, Kone AK, et al. A field trial to assess a blood-stage malaria vaccine. N Engl J Med. 2011;365:1004–13.CrossRefPubMedPubMedCentral
7.
von Bohl A, Kuehn A, Simon N, Ngongang VN, Spehr M, Baumeister S, et al. A WD40-repeat protein unique to malaria parasites associates with adhesion protein complexes and is crucial for blood stage progeny. Malar J. 2015;14:435.CrossRef
8.
Kadekoppala M, Holder AA. Merozoite surface proteins of the malaria parasite: the MSP1 complex and the MSP7 family. Int J Parasitol. 2010;40:1155–61.CrossRefPubMed
9.
Richard D, MacRaild CA, Riglar DT, Chan JA, Foley M, Baum J, et al. Interaction between Plasmodium falciparum apical membrane antigen 1 and the rhoptry neck protein complex defines a key step in the erythrocyte invasion process of malaria parasites. J Biol Chem. 2010;285:14815–22.CrossRefPubMedPubMedCentral
10.
Lin CS, Uboldi AD, Epp C, Bujard H, Tsuboi T, Czabotar PE, et al. Multiple P. falciparum merozoite surface protein 1 complexes mediate merozoite binding to human erythrocytes. J Biol Chem. 2016;291:7703–15.CrossRefPubMed
11.
Kauth CW, Epp C, Bujard H, Lutz R. The merozoite surface protein 1 complex of human malaria parasite Plasmodium falciparum: interactions and arrangements of subunits. J Biol Chem. 2003;278:22257–64.CrossRefPubMed
12.
Cao J, Kaneko O, Thongkukiatkul A, Tachibana M, Otsuki H, Gao Q, et al. Rhoptry neck protein RON2 forms a complex with microneme protein AMA1 in Plasmodium falciparum merozoites. Parasitol Int. 2009;58:29–35.CrossRefPubMed
13.
Cowman AF, Berry D, Baum J. The cellular and molecular basis for malaria parasite invasion of the human red blood cell. J Cell Biol. 2012;198:961–71.CrossRefPubMedPubMedCentral
14.
Ishino T, Chinzei Y, Yuda M. Two proteins with 6-cys motifs are required for malarial parasites to commit to infection of the hepatocyte. Mol Microbiol. 2005;58:1264–75.CrossRefPubMed
15.
van Dijk MR, van Schaijk BC, Khan SM, van Dooren MW, Ramesar J, Kaczanowski S, et al. Three members of the 6-cys protein family of Plasmodium play a role in gamete fertility. PLoS Pathog. 2010;6:e1000853.CrossRefPubMedPubMedCentral
16.
Templeton TJ, Kaslow DC. Identification of additional members define a Plasmodium falciparum gene superfamily which includes Pfs48/45 and Pfs230. Mol Biochem Parasitol. 1999;101:223–7.CrossRefPubMed
17.
Sanders PR, Gilson PR, Cantin GT, Greenbaum DC, Nebl T, Carucci DJ, et al. Distinct protein classes including novel merozoite surface antigens in Raft-like membranes of Plasmodium falciparum. J Biol Chem. 2005;280:40169–76.CrossRefPubMed
18.
Gerloff DL, Creasey A, Maslau S, Carter R. Structural models for the protein family characterized by gamete surface protein Pfs230 of Plasmodium falciparum. Proc Natl Acad Sci USA. 2005;102:13598–603.CrossRefPubMedPubMedCentral
19.
Carter R, Coulson A, Bhatti S, Taylor BJ, Elliott JF. Predicted disulfide-bonded structures for three uniquely related proteins of Plasmodium falciparum, Pfs230, Pfs48/45 and Pf12. Mol Biochem Parasitol. 1995;71:203–10.CrossRefPubMed
20.
Taechalertpaisarn T, Crosnier C, Bartholdson SJ, Hodder AN, Thompson J, Bustamante LY, et al. Biochemical and functional analysis of two Plasmodium falciparum blood-stage 6-cys proteins: P12 and P41. PLoS ONE. 2012;7:e41937.CrossRefPubMedPubMedCentral
21.
van Dijk MR, Douradinha B, Franke-Fayard B, Heussler V, van Dooren MW, van Schaijk B, et al. Genetically attenuated, P36p-deficient malarial sporozoites induce protective immunity and apoptosis of infected liver cells. Proc Natl Acad Sci USA. 2005;102:12194–9.CrossRefPubMedPubMedCentral
22.
23.
VanBuskirk KM, O’Neill MT, De La Vega P, Maier AG, Krzych U, Williams J, et al. Preerythrocytic, live-attenuated Plasmodium falciparum vaccine candidates by design. Proc Natl Acad Sci USA. 2009;106:13004–9.CrossRefPubMedPubMedCentral
24.
Kumar N. Target antigens of malaria transmission blocking immunity exist as a stable membrane bound complex. Parasite Immunol. 1987;9:321–35.CrossRefPubMed
25.
van Dijk MR, Janse CJ, Thompson J, Waters AP, Braks JA, Dodemont HJ, et al. A central role for P48/45 in malaria parasite male gamete fertility. Cell. 2001;104:153–64.CrossRefPubMed
26.
Williamson KC. Pfs230: from malaria transmission-blocking vaccine candidate toward function. Parasite Immunol. 2003;25:351–9.CrossRefPubMed
27.
Read D, Lensen AH, Begarnie S, Haley S, Raza A, Carter R. Transmission-blocking antibodies against multiple, non-variant target epitopes of the Plasmodium falciparum gamete surface antigen Pfs230 are all complement-fixing. Parasite Immunol. 1994;16:511–9.CrossRefPubMed
28.
Quakyi IA, Carter R, Rener J, Kumar N, Good MF, Miller LH. The 230-kDa gamete surface protein of Plasmodium falciparum is also a target for transmission-blocking antibodies. J Immunol. 1987;139:4213–7.PubMed
29.
Tonkin ML, Arredondo SA, Loveless BC, Serpa JJ, Makepeace KA, Sundar N, et al. Structural and biochemical characterization of Plasmodium falciparum 12 (Pf12) reveals a unique interdomain organization and the potential for an antiparallel arrangement with Pf41. J Biol Chem. 2013;288:12805–17.CrossRefPubMedPubMedCentral
30.
Kennedy AT, Schmidt CQ, Thompson JK, Weiss GE, Taechalertpaisarn T, Gilson PR, et al. Recruitment of factor H as a novel complement evasion strategy for blood-stage Plasmodium falciparum infection. J Immunol. 2016;196:1239–48.CrossRefPubMed
31.
32.
Lambros C, Vanderberg JP. Synchronization of Plasmodium falciparum erythrocytic stages in culture. J Parasitol. 1979;65:418–20.CrossRefPubMed
33.
Theisen M, Vuust J, Gottschau A, Jepsen S, Hogh B. Antigenicity and immunogenicity of recombinant glutamate-rich protein of Plasmodium falciparum expressed in Escherichia coli. Clin Diagn Lab Immunol. 1995;2:30–4.PubMedPubMedCentral
34.
Kanodia S, Kumar G, Rizzi L, Pedretti A, Hodder AN, Romeo S, et al. Synthetic peptides derived from the C-terminal 6 kDa region of Plasmodium falciparum SERA5 inhibit the enzyme activity and malaria parasite development. Biochim Biophys Acta. 2014;1840:2765–75.CrossRefPubMed
35.
Ko YP, Liang X, Smith CW, Degen JL, Hook M. Binding of Efb from Staphylococcus aureus to fibrinogen blocks neutrophil adherence. J Biol Chem. 2011;286:9865–74.CrossRefPubMedPubMedCentral
36.
Wu Y, Li Q, Chen XZ. Detecting protein–protein interactions by Far western blotting. Nat Protoc. 2007;2:3278–84.CrossRefPubMed
37.
Wickramarachchi T, Devi YS, Mohmmed A, Chauhan VS. Identification and characterization of a novel Plasmodium falciparum merozoite apical protein involved in erythrocyte binding and invasion. PLoS ONE. 2008;3:e1732.CrossRefPubMedPubMedCentral
38.
Courtin D, Oesterholt M, Huismans H, Kusi K, Milet J, Badaut C, et al. The quantity and quality of African children’s IgG responses to merozoite surface antigens reflect protection against Plasmodium falciparum malaria. PLoS ONE. 2009;4:e7590.CrossRefPubMedPubMedCentral
39.
Tarique M, Satsangi AT, Ahmad M, Singh S, Tuteja R. Plasmodium falciparum MLH is schizont stage specific endonuclease. Mol Biochem Parasitol. 2012;181:153–61.CrossRefPubMed
40.
Singh S, Pandey K, Chattopadhayay R, Yazdani SS, Lynn A, Bharadwaj A, et al. Biochemical, biophysical, and functional characterization of bacterially expressed and refolded receptor binding domain of Plasmodium vivax duffy-binding protein. J Biol Chem. 2001;276:17111–6.CrossRefPubMed
41.
Ramasamy G, Gupta D, Mohmmed A, Chauhan VS. Characterization and localization of Plasmodium falciparum homolog of prokaryotic ClpQ/HslV protease. Mol Biochem Parasitol. 2007;152:139–48.CrossRefPubMed
42.
Garcia J, Curtidor H, Pinzon CG, Vanegas M, Moreno A, Patarroyo ME. Identification of conserved erythrocyte binding regions in members of the Plasmodium falciparum Cys6 lipid raft-associated protein family. Vaccine. 2009;27:3953–62.CrossRefPubMed
43.
Baldwin MR, Li X, Hanada T, Liu SC, Chishti AH. Merozoite surface protein 1 recognition of host glycophorin A mediates malaria parasite invasion of red blood cells. Blood. 2015;125:2704–11.CrossRefPubMedPubMedCentral
44.
Williamson KC, Keister DB, Muratova O, Kaslow DC. Recombinant Pfs230, a Plasmodium falciparum gametocyte protein, induces antisera that reduce the infectivity of Plasmodium falciparum to mosquitoes. Mol Biochem Parasitol. 1995;75:33–42.CrossRefPubMed
45.
Arredondo SA, Cai M, Takayama Y, MacDonald NJ, Anderson DE, Aravind L, et al. Structure of the Plasmodium 6-cysteine s48/45 domain. Proc Natl Acad Sci USA. 2012;109:6692–7.CrossRefPubMedPubMedCentral
46.
Hostetler JB, Sharma S, Bartholdson SJ, Wright GJ, Fairhurst RM, Rayner JC. A library of Plasmodium vivax recombinant merozoite proteins reveals new vaccine candidates and protein-protein interactions. PLoS Negl Trop Dis. 2015;9:e0004264.CrossRefPubMedPubMedCentral
47.
Ranjan R, Chugh M, Kumar S, Singh S, Kanodia S, Hossain MJ, et al. Proteome analysis reveals a large merozoite surface protein-1 associated complex on the Plasmodium falciparum merozoite surface. J Proteom Res. 2011;10:680–91.CrossRef
48.
Li G, Basagoudanavar SH, Gowda DC. Effect of GPI anchor moiety on the immunogenicity of DNA plasmids encoding the 19-kDa C-terminal portion of Plasmodium falciparum MSP-1. Parasite Immunol. 2008;30:315–22.CrossRefPubMedPubMedCentral
49.
Feller T, Thom P, Koch N, Spiegel H, Addai-Mensah O, Fischer R, et al. Plant-based production of recombinant Plasmodium surface protein pf38 and evaluation of its potential as a vaccine candidate. PLoS ONE. 2013;8:e79920.CrossRefPubMedPubMedCentral
50.
Srinivasan P, Ekanem E, Diouf A, Tonkin ML, Miura K, Boulanger MJ, et al. Immunization with a functional protein complex required for erythrocyte invasion protects against lethal malaria. Proc Natl Acad Sci USA. 2014;111:10311–6.CrossRefPubMedPubMedCentral
51.
Dziegiel M, Borre MB, Jepsen S, Hogh B, Petersen E, Vuust J. Recombinant Plasmodium falciparum glutamate rich protein; purification and use in enzyme-linked immunosorbent assay. Am J Trop Med Hyg. 1991;44:306–13.PubMed
52.
Dziegiel M, Rowe P, Bennett S, Allen SJ, Olerup O, Gottschau A, et al. Immunoglobulin M and G antibody responses to Plasmodium falciparum glutamate-rich protein: correlation with clinical immunity in Gambian children. Infect Immun. 1993;61:103–8.PubMedPubMedCentral
Metadata
Title
A novel Pfs38 protein complex on the surface of Plasmodium falciparum blood-stage merozoites
Authors
Gourab Paul
Arunaditya Deshmukh
Inderjeet Kaur
Sumit Rathore
Surbhi Dabral
Ashutosh Panda
Susheel Kumar Singh
Asif Mohmmed
Michael Theisen
Pawan Malhotra
Publication date
01-12-2017
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2017
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/s12936-017-1716-0

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