Fractionation and identification of proteins by 2-DE and MS: towards a proteomic analysis of Plasmodium falciparum.
Since completion of genome sequencing of the malarial parasite Plasmodium falciparum, proteomic tools for the identification of parasite proteins have become particularly attractive as they allow a more thorough interpretation of these data. Recent advances in 2-D PAGE, MS, and bioinformatics have created great opportunities for mapping and characterization of protein populations. We employed these improvements in a proteomic approach for the analysis of proteins detected in two blood stages of P. falciparum, (i) in the schizont stage and (ii) in the merozoite stage. For the isolation of merozoites, we introduced a new protocol based on the preparation of clustered structures of merozoites upon treatment of cultures with the common cysteine proteinase inhibitor E64. Peptide mass fingerprints of excised and trypsinated protein spots, acquired by MALDI-TOF MS were generated to identify a variety of proteins. Moreover, prefractionation procedures were used to enrich and map low-abundance proteins in protein samples. The data demonstrate that classic proteomic analyses using 2-D PAGE are now feasible for P. falciparum and represent the first step in the direction of creating 2-D reference maps for this medically most relevant protozoon.
Gelhaus
Christoph
C
Research Center for Infectious Diseases of the University of Würzburg, Würzburg, Germany. cgelhaus@zoologie.uni-kiel.de
Fritsch
Jürgen
J
Krause
Eberhard
E
Leippe
Matthias
M
D000818:Animals; D002478:Cells, Cultured; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D004912:Erythrocytes / Q000166:cytology / Q000469:parasitology; D006801:Humans; D066298:In Vitro Techniques; D010963:Plasmodium falciparum / Q000166:cytology / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D015800:Protozoan Proteins / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D015853:Cysteine Proteinase Inhibitors; D020543:Proteome; D015800:Protozoan Proteins