Proteomic profiling of Plasmodium falciparum through improved, semi-quantitative two-dimensional gel electrophoresis

dc.contributor.authorSmit, Salome
dc.contributor.authorStoychev, Stoyan H.
dc.contributor.authorLouw, Abraham Izak
dc.contributor.authorBirkholtz, Lyn-Marie
dc.contributor.emaillbirkholtz@up.ac.zaen_US
dc.date.accessioned2011-02-22T09:38:17Z
dc.date.available2011-02-22T09:38:17Z
dc.date.issued2010-03
dc.description.abstractTwo-dimensional gel electrophoresis (2-DE) is one of the most commonly used technologies to obtain a snapshot of the proteome at any specific time. However, its application to study the Plasmodial (malaria parasite) proteome is still limited due to inefficient extraction and detection methods and the extraordinarily large size of some proteins. Here, we report an optimized protein extraction method, the most appropriate methods for Plasmodial protein quantification and 2-DE detection, and finally protein identification by mass spectrometry (MS). Linear detection of Plasmodial proteins in a optimized lysis buffer was only possible with the 2-D Quant kit, and of the four stains investigated, Flamingo Pink was superior regarding sensitivity, linearity, and excellent MS-compatibility. 2-DE analyses of the Plasmodial proteome using this methodology resulted in the reliable detection of 349 spots and a 95% success rate in MS/MS identification. Subsequent application to the analyses of the Plasmodial ring and trophozoite proteomes ultimately resulted in the identification of 125 protein spots, which constituted 57 and 49 proteins from the Plasmodial ring and trophozoite stages, respectively. This study additionally highlights the presence of various isoforms within the Plasmodial proteome, which is of significant biological importance within the Plasmodial parasite during development in the intraerythrocytic developmental cycle.en
dc.description.sponsorshipFunding was provided by the National Research Foundation (NRF Grant FA2004051300055, Thuthuka TTK2006061500031 and Prestigious Bursary to S.S.), the South African Malaria Initiative (www.sami.org.za) and the University of Pretoria.en_US
dc.identifier.citationSmit, S, Stoychev, S, Louw, AI & Birkholtz, LM 2010, 'Proteomic profiling of Plasmodium falciparum through improved, semi-quantitative two-dimensional gel electrophoresis', Journal of Proteome Research, vol. 9, no. 5, pp. 2170-2181. [http://pubs.acs.org/journal/jprobs]en
dc.identifier.issn1535-3893
dc.identifier.issn1535-3907 (online)
dc.identifier.other10.1021/pr9009244
dc.identifier.urihttp://hdl.handle.net/2263/15916
dc.language.isoenen_US
dc.publisherAmerican Chemical Societyen_US
dc.rights© 2010 American Chemical Society. This document is the Accepted Manuscript version of a Published Work that appeared in final form in Journal of Proteome Research, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://pubs.acs.org/journal/jprobs.en_US
dc.subject2-D gel electrophoresisen
dc.subjectProtein extractionen
dc.subjectIsoformsen
dc.subject.lcshPlasmodium falciparumen
dc.subject.lcshMalariaen
dc.subject.lcshProteomicsen
dc.subject.lcshGel electrophoresisen
dc.titleProteomic profiling of Plasmodium falciparum through improved, semi-quantitative two-dimensional gel electrophoresisen
dc.typePostprint Articleen

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