Journal
MOLECULAR AND BIOCHEMICAL PARASITOLOGY
Volume 136, Issue 1, Pages 51-62Publisher
ELSEVIER
DOI: 10.1016/j.molbiopara.2004.02.009
Keywords
Leishmania mexicana; differentiation; proteomics; 2D gel electrophoresis
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Funding
- Wellcome Trust [061343] Funding Source: Medline
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We have resolved the proteome of axenically differentiated Leishmania mexicana parasites by two-dimensional gel electrophoresis (2DE), employing optimised, robust and reproducible procedures, and visualised (by silver staining) approximately 2000 protein species in each of three developmental stages: procyclic promastigotes, metacyclic promastigotes and amastigotes. This analysis has used homogeneous populations of these parasite stages, characterised according to their morphology, protease and nuclease activity profiles and expression of stage-specific antigens. Following comparison of the whole proteome profiles between stages, 47 spots were found to be stage-specific, while a further 100 spots changed in intensity during differentiation. The majority of unique spots were expressed during the infective stages of parasite differentiation, metacyclic promastigotes and amastigotes. CapLC-QTOF mass spectrometry has allowed the identification of 47 protein species to date, including a number which are only detected in the amastigote stage. Proteins identified are members of eight functionally related groupings, some of which are implicated in infectivity and host-parasite interactions. (C) 2004 Elsevier B.V. All rights reserved.
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