J Proteomics. 2014 Mar 21. pii: S1874-3919(14)00130-4. doi: 10.1016/j.jprot.2014.03.008. [Epub ahead of print]

Proteomic characterization of the subpellicular cytoskeleton of Toxoplasma gondii tachyzoites

Gómez de León CT¹, Martín RD¹, Hernández GM², Pozos SG³, Ambrosio JR⁴, Flores RM⁵.

 

Toxoplasma, the causative agent of toxoplasmosis in animals and humans, has a subpellicular cytoskeleton that is involved in motility, cell shape and invasion. Knowledge of components of the cytoskeleton is necessary to understand the invasion mechanisms as well as for the identification of possible therapeutic targets. To date, most cytoskeletal components of Toxoplasma remain unidentified due mainly to the lack of reproducible methods for their isolation. Based on the successful isolation of the cytoskeleton, it was possible to report by the first time, the proteomic characterization of the subpellicular cytoskeleton of Toxoplasma formed by 95 cytoskeletal proteins through proteomic analysis by tandem mass spectrometry of one dimension SDS PAGE. By bioinformatic analysis of the data, proteins were classified as: Eighteen conventional cytoskeletal proteins; 10 inner membrane complex proteins, included 7 with alveolin repeats; 5 new proteins with alveolin like repeats; 37 proteins associated with other organelles and 25 novel proteins of unknown function. One of the alveolin like protein not previously described in Toxoplasma named as TgArticulin was partially characterized with a specific monoclonal antibody. Presence of TgArticulin was exclusively associated to the cytoskeleton fraction with a cortical distribution. Functions for the several molecules identified are proposed. This article is part of a Special Issue entitled: Proteomics, mass spectrometry and peptidomics, Cancun 2013. Industrial significance This manuscript describes, for the first time, the proteome of the subpellicular cytoskeleton of Toxoplasma gondii. The importance of this study is related to the role of the cytoskeleton in the highly invasive capability of a parasite that causes abortion, blindness, and death by encephalitis in immunocompromised patients. Proteomic characterization of the cytoskeleton of T. gondii tachyzoites was possible by the development of a successful procedure for the isolation of the subpellicular cytoskeleton. Knowledge of the composition of the cytoskeleton of Toxoplasma is fundamental for the understanding of the motility and host cell invasion mechanisms, and for the future design and development of toxoplasmicidal drugs with effects against specific components of the cytoskeleton of this parasite that are absent in mammal host cells.
Copyright © 2014 Elsevier B.V. All rights reserved.

KEYWORDS:

Toxoplasma, articulin, proteome, subpellicular cytoskeleton, tandem mass spectrometry

PMID:

24662527

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