A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyte
Introduction: We propose a new method for the selective labeling, isolation and electrophoretic analysis of the Plasmodium falciparum protein exposed on the erythrocyte cell surface. Historically, membrane surface proteins have been isolated using a surface biotinylation followed by capture of biot...
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The Journal of Infection in Developing Countries
2012-01-01
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| Series: | Journal of Infection in Developing Countries |
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| Online Access: | https://jidc.org/index.php/journal/article/view/2386 |
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| author | Emanuela Ferru Anntonella Pantaleo Francesco Turrini |
| author_facet | Emanuela Ferru Anntonella Pantaleo Francesco Turrini |
| author_sort | Emanuela Ferru |
| collection | DOAJ |
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Introduction: We propose a new method for the selective labeling, isolation and electrophoretic analysis of the Plasmodium falciparum protein exposed on the erythrocyte cell surface. Historically, membrane surface proteins have been isolated using a surface biotinylation followed by capture of biotin-conjugated protein via an avidin/streptavidin-coated solid support. The major drawback of the standard methods has been the labeling of internal proteins due to fast internalization of biotin.
Methodology: To solve this problem, we used a biotin label that does not permeate through the membrane. As a further precaution to avoid the purification of non surface exposed proteins, we directly challenged whole labeled cells with avidin coated beads and then solubilized them using non ionic detergents.
Results: A marked enrichment of most of the RBC membrane proteins known to face the external surface of the membrane validated the specificity of the method; furthermore, only small amounts of haemoglobin and cytoskeletal proteins were detected. A wide range of P. falciparum proteins were additionally described to be exposed on the erythrocyte surface. Some of them have been previously observed and used as vaccine candidates while a number of newly described antigens have been presently identified. Those antigens require further characterization and validation with additional methods.
Conclusion: Surface proteins preparations were very reproducible and identification of proteins by mass spectrometry has been demonstrated to be feasible and effective.
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| format | Article |
| id | doaj-art-4e197df39e5e49829670f4bef9a36e15 |
| institution | OA Journals |
| issn | 1972-2680 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | The Journal of Infection in Developing Countries |
| record_format | Article |
| series | Journal of Infection in Developing Countries |
| spelling | doaj-art-4e197df39e5e49829670f4bef9a36e152025-08-20T02:16:06ZengThe Journal of Infection in Developing CountriesJournal of Infection in Developing Countries1972-26802012-01-0160610.3855/jidc.2386A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyteEmanuela Ferru0Anntonella Pantaleo1Francesco Turrini2University of Turin, 10126 Turin, ItalyUniversity of Sassari, 07100, ItalyUniversity of Turin, 10126 Turin, Italy Introduction: We propose a new method for the selective labeling, isolation and electrophoretic analysis of the Plasmodium falciparum protein exposed on the erythrocyte cell surface. Historically, membrane surface proteins have been isolated using a surface biotinylation followed by capture of biotin-conjugated protein via an avidin/streptavidin-coated solid support. The major drawback of the standard methods has been the labeling of internal proteins due to fast internalization of biotin. Methodology: To solve this problem, we used a biotin label that does not permeate through the membrane. As a further precaution to avoid the purification of non surface exposed proteins, we directly challenged whole labeled cells with avidin coated beads and then solubilized them using non ionic detergents. Results: A marked enrichment of most of the RBC membrane proteins known to face the external surface of the membrane validated the specificity of the method; furthermore, only small amounts of haemoglobin and cytoskeletal proteins were detected. A wide range of P. falciparum proteins were additionally described to be exposed on the erythrocyte surface. Some of them have been previously observed and used as vaccine candidates while a number of newly described antigens have been presently identified. Those antigens require further characterization and validation with additional methods. Conclusion: Surface proteins preparations were very reproducible and identification of proteins by mass spectrometry has been demonstrated to be feasible and effective. https://jidc.org/index.php/journal/article/view/2386Plasmodium falciparumerythrocytemalariasurface protein |
| spellingShingle | Emanuela Ferru Anntonella Pantaleo Francesco Turrini A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyte Journal of Infection in Developing Countries Plasmodium falciparum erythrocyte malaria surface protein |
| title | A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyte |
| title_full | A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyte |
| title_fullStr | A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyte |
| title_full_unstemmed | A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyte |
| title_short | A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyte |
| title_sort | new method for the capture of surface proteins in plasmodium falciparum parasitized erythrocyte |
| topic | Plasmodium falciparum erythrocyte malaria surface protein |
| url | https://jidc.org/index.php/journal/article/view/2386 |
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