Molecular Characterization and Immuno-Reactivity Patterns of a Novel Plasmodium falciparum Armadillo-Type Repeat Protein, PfATRP
Nearly half of the genes in the Plasmodium falciparum genome have not yet been functionally investigated. We used homology-based structural modeling to identify multiple copies of Armadillo repeats within one uncharacterized gene expressed during the intraerythrocytic stages, PF3D7_0410600, subseque...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-03-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fcimb.2020.00114/full |
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| author | Emmanuel Amlabu Emmanuel Amlabu Philip Ilani Grace Opoku Prince B. Nyarko Evelyn Quansah Laty G. Thiam Manfred Anim Reuben Ayivor-Djanie Reuben Ayivor-Djanie Ojo-ajogu Akuh Henrietta Mensah-Brown Julian C. Rayner Gordon A. Awandare Gordon A. Awandare |
| author_facet | Emmanuel Amlabu Emmanuel Amlabu Philip Ilani Grace Opoku Prince B. Nyarko Evelyn Quansah Laty G. Thiam Manfred Anim Reuben Ayivor-Djanie Reuben Ayivor-Djanie Ojo-ajogu Akuh Henrietta Mensah-Brown Julian C. Rayner Gordon A. Awandare Gordon A. Awandare |
| author_sort | Emmanuel Amlabu |
| collection | DOAJ |
| description | Nearly half of the genes in the Plasmodium falciparum genome have not yet been functionally investigated. We used homology-based structural modeling to identify multiple copies of Armadillo repeats within one uncharacterized gene expressed during the intraerythrocytic stages, PF3D7_0410600, subsequently referred to as P. falciparum Armadillo-Type Repeat Protein (PfATRP). Soluble recombinant PfATRP was expressed in a bacterial expression system, purified to apparent homogeneity and the identity of the recombinant PfATRP was confirmed by mass spectrometry. Affinity-purified α-PfATRP rabbit antibodies specifically recognized the recombinant protein. Immunofluorescence assays revealed that α-PfATRP rabbit antibodies reacted with P. falciparum schizonts. Anti-PfATRP antibody exhibited peripheral staining patterns around the merozoites. Given the localization of PfATRP in merozoites, we tested for an egress phenotype during schizont arrest assays and demonstrated that native PfATRP is inaccessible on the surface of merozoites in intact schizonts. Dual immunofluorescence assays with markers for the inner membrane complex (IMC) and microtubules suggest partial colocalization in both asexual and sexual stage parasites. Using the soluble recombinant PfATRP in a screen of plasma samples revealed that malaria-infected children have naturally acquired PfATRP-specific antibodies. |
| first_indexed | 2024-12-13T01:57:31Z |
| format | Article |
| id | doaj.art-37e7c64db27a4e3f91e6e2e225d6a275 |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-12-13T01:57:31Z |
| publishDate | 2020-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-37e7c64db27a4e3f91e6e2e225d6a2752022-12-22T00:03:21ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882020-03-011010.3389/fcimb.2020.00114507457Molecular Characterization and Immuno-Reactivity Patterns of a Novel Plasmodium falciparum Armadillo-Type Repeat Protein, PfATRPEmmanuel Amlabu0Emmanuel Amlabu1Philip Ilani2Grace Opoku3Prince B. Nyarko4Evelyn Quansah5Laty G. Thiam6Manfred Anim7Reuben Ayivor-Djanie8Reuben Ayivor-Djanie9Ojo-ajogu Akuh10Henrietta Mensah-Brown11Julian C. Rayner12Gordon A. Awandare13Gordon A. Awandare14West African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaDepartment of Biochemistry, Kogi State University, Anyigba, NigeriaWest African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaWest African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaWest African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaWest African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaWest African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaWest African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaWest African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaDepartment of Biomedical Sciences, SBBS, University of Health and Allied Sciences, Ho, GhanaWest African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaWest African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaCambridge Institute for Medical Research, University of Cambridge, Cambridge, United KingdomWest African Center for Cell Biology of Infectious Pathogens, University of Ghana, Accra, GhanaDepartment of Biochemistry, Cell and Molecular Biology, College of Basic and Applied Sciences, University of Ghana, Accra, GhanaNearly half of the genes in the Plasmodium falciparum genome have not yet been functionally investigated. We used homology-based structural modeling to identify multiple copies of Armadillo repeats within one uncharacterized gene expressed during the intraerythrocytic stages, PF3D7_0410600, subsequently referred to as P. falciparum Armadillo-Type Repeat Protein (PfATRP). Soluble recombinant PfATRP was expressed in a bacterial expression system, purified to apparent homogeneity and the identity of the recombinant PfATRP was confirmed by mass spectrometry. Affinity-purified α-PfATRP rabbit antibodies specifically recognized the recombinant protein. Immunofluorescence assays revealed that α-PfATRP rabbit antibodies reacted with P. falciparum schizonts. Anti-PfATRP antibody exhibited peripheral staining patterns around the merozoites. Given the localization of PfATRP in merozoites, we tested for an egress phenotype during schizont arrest assays and demonstrated that native PfATRP is inaccessible on the surface of merozoites in intact schizonts. Dual immunofluorescence assays with markers for the inner membrane complex (IMC) and microtubules suggest partial colocalization in both asexual and sexual stage parasites. Using the soluble recombinant PfATRP in a screen of plasma samples revealed that malaria-infected children have naturally acquired PfATRP-specific antibodies.https://www.frontiersin.org/article/10.3389/fcimb.2020.00114/fullPfATRPrecombinant proteinimmunolocalizationimmunoreactivityserosurveilance |
| spellingShingle | Emmanuel Amlabu Emmanuel Amlabu Philip Ilani Grace Opoku Prince B. Nyarko Evelyn Quansah Laty G. Thiam Manfred Anim Reuben Ayivor-Djanie Reuben Ayivor-Djanie Ojo-ajogu Akuh Henrietta Mensah-Brown Julian C. Rayner Gordon A. Awandare Gordon A. Awandare Molecular Characterization and Immuno-Reactivity Patterns of a Novel Plasmodium falciparum Armadillo-Type Repeat Protein, PfATRP Frontiers in Cellular and Infection Microbiology PfATRP recombinant protein immunolocalization immunoreactivity serosurveilance |
| title | Molecular Characterization and Immuno-Reactivity Patterns of a Novel Plasmodium falciparum Armadillo-Type Repeat Protein, PfATRP |
| title_full | Molecular Characterization and Immuno-Reactivity Patterns of a Novel Plasmodium falciparum Armadillo-Type Repeat Protein, PfATRP |
| title_fullStr | Molecular Characterization and Immuno-Reactivity Patterns of a Novel Plasmodium falciparum Armadillo-Type Repeat Protein, PfATRP |
| title_full_unstemmed | Molecular Characterization and Immuno-Reactivity Patterns of a Novel Plasmodium falciparum Armadillo-Type Repeat Protein, PfATRP |
| title_short | Molecular Characterization and Immuno-Reactivity Patterns of a Novel Plasmodium falciparum Armadillo-Type Repeat Protein, PfATRP |
| title_sort | molecular characterization and immuno reactivity patterns of a novel plasmodium falciparum armadillo type repeat protein pfatrp |
| topic | PfATRP recombinant protein immunolocalization immunoreactivity serosurveilance |
| url | https://www.frontiersin.org/article/10.3389/fcimb.2020.00114/full |
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