Proteomic analysis of Leishmania mexicana differentiation.
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.
Nugent
Philip G
PG
Wellcome Trust Laboratories for Molecular Parasitology, Department of Biological Sciences, Centre for Molecular Microbiology and Infection, Imperial College London, Exhibition Road, London SW7 2AZ, UK.
Karsani
Saiful A
SA
Wait
Robin
R
Tempero
Jane
J
Smith
Deborah F
DF
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D007894:Leishmania mexicana / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D008018:Life Cycle Stages; D013058:Mass Spectrometry; D020543:Proteome; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016622:Silver Staining
D020543:Proteome; D015800:Protozoan Proteins