Analysis of Stage-Specific Expression of Basic Proteins in Leishmania infantum

作者：Brotherton Marie Christine; Racine Gina; Foucher Aude L; Drummelsmith Jolyne; Papadopoulou Barbara; Ouellette Marc^*

来源：Journal of Proteome Research, 2010, 9(8): 3842-3853.

DOI：10.1021/pr100048m

• Leishmania infantum
• free-flow electrophoresis (FFE)
• basic proteins
• two-dimensional gel electrophoresis
• cytodifferentiation
• elongation factor 1-alpha

Prior analyses of the proteome of the protozoan parasite Leishmania have underrepresented basic proteins. Here, we applied protein fractionation by isoelectric point (pI) using free-flow electrophoresis (FFE) to study stage-specific expression of basic proteins in this pathogen. Overall, we resolved 2469 protein spots in both the flagellated promastigote and the nonmotile amastigote forms in the basic range by two-dimensional gel electrophoresis (2-DE). Highly basic proteins were enriched by FFE fractionation, allowing many to be identified and characterized for the first time by proteomics analysis. Among proteins upregulated in the promastigote stage, we found glycolytic enzymes and flagellar proteins. Proteins upregulated in the amastigote stage included enzymes involved in gluconeogenesis and fatty acid beta-oxidation. In both life stages, many proteins were found in multiple spots or as proteolytic fragments, suggesting that extensive post-translational modification and processing occur. Interestingly, evidence was obtained suggesting that some of these processes may be stage-specific.

• 出版日期2010-8