GPI- Anchored Proteomes and Their Biochemical Pathway in Alveolates: Evolutionary Insights and Heterologous Expression in Tetrahymena Thermophila

Glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) play essential roles in eukaryotic cell biology, yet their evolutionary diversity across alveolates remains poorly understood. Here, we conducted a comparative genomic analysis of GPI biosynthetic pathways and GPI-AP repertoires across r...

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Main Authors: Puca, Gervasio, Montes, Maria G., Orlowski, Juan, Tomazic, Mariela Luján, Gonzalez Maglio, Daniel H., de la Fuente, Carolina, Florin-Christensen, Mónica, Schnittger, Leonhard, Nusblat, Alejandro D.
Format: info:ar-repo/semantics/artículo
Language:Inglés
Published: Wiley 2025
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/24811
https://onlinelibrary.wiley.com/doi/10.1111/jeu.70055
https://doi.org/10.1111/jeu.70055
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author Puca, Gervasio
Montes, Maria G.
Orlowski, Juan
Tomazic, Mariela Luján
Gonzalez Maglio, Daniel H.
de la Fuente, Carolina
Florin-Christensen, Mónica
Schnittger, Leonhard
Nusblat, Alejandro D.
author_browse Florin-Christensen, Mónica
Gonzalez Maglio, Daniel H.
Montes, Maria G.
Nusblat, Alejandro D.
Orlowski, Juan
Puca, Gervasio
Schnittger, Leonhard
Tomazic, Mariela Luján
de la Fuente, Carolina
author_facet Puca, Gervasio
Montes, Maria G.
Orlowski, Juan
Tomazic, Mariela Luján
Gonzalez Maglio, Daniel H.
de la Fuente, Carolina
Florin-Christensen, Mónica
Schnittger, Leonhard
Nusblat, Alejandro D.
author_sort Puca, Gervasio
collection INTA Digital
description Glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) play essential roles in eukaryotic cell biology, yet their evolutionary diversity across alveolates remains poorly understood. Here, we conducted a comparative genomic analysis of GPI biosynthetic pathways and GPI-AP repertoires across representative species of Ciliophora, Apicomplexa, and Dinoflagellata. Our findings reveal lineage-specific patterns of gene loss, expansion, and functional divergence, with ciliates exhibiting reduced canonical enzyme sets but expanded trafficking components. Using a benchmarked pipeline of GPI-AP predictors combined with signal peptide detection, we identified candidate GPI-anchored proteins and performed functional annotation to determine enriched Gene Ontology terms. To complement the bioinformatic analyses, we expressed the apicomplexan GPI-anchored antigen GPI4 from Babesia bovis in Tetrahymena thermophila. This approach extends previous efforts that used Tetrahymena as a heterologous expression system for parasite surface antigens. The recombinant protein localized to ciliary membranes and showed features consistent with proper processing. This study highlights the evolutionary plasticity of GPI systems in alveolates and reinforces the utility of Tetrahymena as a eukaryotic platform for the production and analysis of GPI-anchored proteins with potential diagnostic or immunoprophylactic applications.
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spelling INTA248112025-12-30T12:49:15Z GPI- Anchored Proteomes and Their Biochemical Pathway in Alveolates: Evolutionary Insights and Heterologous Expression in Tetrahymena Thermophila Puca, Gervasio Montes, Maria G. Orlowski, Juan Tomazic, Mariela Luján Gonzalez Maglio, Daniel H. de la Fuente, Carolina Florin-Christensen, Mónica Schnittger, Leonhard Nusblat, Alejandro D. Tetrahymena Célula Eucariotica Citología Estructura Genética Eukaryotic Cells Cytology Genetic Structures Anchored Proteomes Glycosylphosphatidylinositol Proteomas Anclados Glicosilfosfatidilinositol Tetrahymena thermophila Glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) play essential roles in eukaryotic cell biology, yet their evolutionary diversity across alveolates remains poorly understood. Here, we conducted a comparative genomic analysis of GPI biosynthetic pathways and GPI-AP repertoires across representative species of Ciliophora, Apicomplexa, and Dinoflagellata. Our findings reveal lineage-specific patterns of gene loss, expansion, and functional divergence, with ciliates exhibiting reduced canonical enzyme sets but expanded trafficking components. Using a benchmarked pipeline of GPI-AP predictors combined with signal peptide detection, we identified candidate GPI-anchored proteins and performed functional annotation to determine enriched Gene Ontology terms. To complement the bioinformatic analyses, we expressed the apicomplexan GPI-anchored antigen GPI4 from Babesia bovis in Tetrahymena thermophila. This approach extends previous efforts that used Tetrahymena as a heterologous expression system for parasite surface antigens. The recombinant protein localized to ciliary membranes and showed features consistent with proper processing. This study highlights the evolutionary plasticity of GPI systems in alveolates and reinforces the utility of Tetrahymena as a eukaryotic platform for the production and analysis of GPI-anchored proteins with potential diagnostic or immunoprophylactic applications. Instituto de Patobiología Fil: Puca, Gervasio. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Nanobiotecnología (NANOBIOTEC); Argentina Fil; Puca, Gervasio. Universidad de Buenos Aires (UBA). Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología (NANOBIOTEC); Argentina Fil: Montes, Maria G. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Nanobiotecnología (NANOBIOTEC); Argentina Fil: Montes, Maria G. Universidad de Buenos Aires (UBA). Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología (NANOBIOTEC); Argentina Fil: Orlowski, Juan. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Nanobiotecnología (NANOBIOTEC); Argentina Fil: Orlowski, Juan. Universidad de Buenos Aires (UBA). Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología (NANOBIOTEC); Argentina Fil: Tomazic, Mariela Luján. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina Fil: Tomazic, Mariela Luján. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina Fil: Gonzalez Maglio, Daniel H. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni (IDEHU); Argentina Fil: Gonzalez Maglio, Daniel H. Universidad de Buenos Aires (UBA). Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni (IDEHU); Argentina Fil: de la Fuente, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Nanobiotecnología (NANOBIOTEC); Argentina Fil: de la Fuente, Carolina. Universidad de Buenos Aires (UBA). Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología (NANOBIOTEC); Argentina Fil: Florin-Christensen, Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina Fil: Florin-Christensen, Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina Fil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina Fil: Schnittger, Leonhard. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina Fil: Nusblat, Alejandro D. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Nanobiotecnología (NANOBIOTEC); Argentina Fil: Nusblat, Alejandro D. Universidad de Buenos Aires (UBA). Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología (NANOBIOTEC); Argentina 2025-12-30T12:29:39Z 2025-12-30T12:29:39Z 2025-11 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/24811 https://onlinelibrary.wiley.com/doi/10.1111/jeu.70055 1550-7408 1066-5234 https://doi.org/10.1111/jeu.70055 eng info:eu-repo/semantics/restrictedAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Wiley Journal of Eukaryotic Microbiology 73 (1) : e70055 (January/February 2026)
spellingShingle Tetrahymena
Célula Eucariotica
Citología
Estructura Genética
Eukaryotic Cells
Cytology
Genetic Structures
Anchored Proteomes
Glycosylphosphatidylinositol
Proteomas Anclados
Glicosilfosfatidilinositol
Tetrahymena thermophila
Puca, Gervasio
Montes, Maria G.
Orlowski, Juan
Tomazic, Mariela Luján
Gonzalez Maglio, Daniel H.
de la Fuente, Carolina
Florin-Christensen, Mónica
Schnittger, Leonhard
Nusblat, Alejandro D.
GPI- Anchored Proteomes and Their Biochemical Pathway in Alveolates: Evolutionary Insights and Heterologous Expression in Tetrahymena Thermophila
title GPI- Anchored Proteomes and Their Biochemical Pathway in Alveolates: Evolutionary Insights and Heterologous Expression in Tetrahymena Thermophila
title_full GPI- Anchored Proteomes and Their Biochemical Pathway in Alveolates: Evolutionary Insights and Heterologous Expression in Tetrahymena Thermophila
title_fullStr GPI- Anchored Proteomes and Their Biochemical Pathway in Alveolates: Evolutionary Insights and Heterologous Expression in Tetrahymena Thermophila
title_full_unstemmed GPI- Anchored Proteomes and Their Biochemical Pathway in Alveolates: Evolutionary Insights and Heterologous Expression in Tetrahymena Thermophila
title_short GPI- Anchored Proteomes and Their Biochemical Pathway in Alveolates: Evolutionary Insights and Heterologous Expression in Tetrahymena Thermophila
title_sort gpi anchored proteomes and their biochemical pathway in alveolates evolutionary insights and heterologous expression in tetrahymena thermophila
topic Tetrahymena
Célula Eucariotica
Citología
Estructura Genética
Eukaryotic Cells
Cytology
Genetic Structures
Anchored Proteomes
Glycosylphosphatidylinositol
Proteomas Anclados
Glicosilfosfatidilinositol
Tetrahymena thermophila
url http://hdl.handle.net/20.500.12123/24811
https://onlinelibrary.wiley.com/doi/10.1111/jeu.70055
https://doi.org/10.1111/jeu.70055
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