Comparative proteomic analysis of metabolically labelled proteins from Plasmodium falciparum isolates with different adhesion properties.
The virulence of Plasmodium falciparum relates in part to the cytoadhesion characteristics of parasitized erythrocytes but the molecular basis of the different qualitative and quantitative binding phenotypes is incompletely understood. This paucity of information is due partly to the difficulty in working with membrane proteins, the variant nature of these surface antigens and their relatively low abundance. To address this two-dimensional (2D) protein profiles of closely related, but phenotypically different laboratory strains of P. falciparum have been characterized using proteomic approaches. Since the mature erythrocyte has no nucleus and no protein synthesis capability, metabolic labelling of proteins was used to selectively identify parasite proteins and increase detection sensitivity.A small number of changes (less than 10) were observed between four different P. falciparum laboratory strains with distinctive cytoadherence properties using metabolic labelling, with more parasite protein changes found in trophozoite iRBCs than ring stage. The combination of metabolic labelling and autoradiography can therefore be used to identify parasite protein differences, including quantitative ones, and in some cases to obtain protein identifications by mass spectrometry. The results support the suggestion that the membrane protein profile may be related to cytoadherent properties of the iRBCs. Most changes between parasite variants were differences in iso-electric point indicating differential protein modification rather than the presence or absence of a specific peptide.
Wu
Yang
Y
Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK. ywu@liv.ac.uk
Craig
Alister
A
D000818:Animals; D002448:Cell Adhesion; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004910:Erythrocyte Membrane / Q000737:chemistry* / Q000469:parasitology; D006790:Host-Parasite Interactions / Q000502:physiology; D006801:Humans; D010963:Plasmodium falciparum / Q000737:chemistry* / Q000502:physiology; D021381:Protein Transport; D020543:Proteome / Q000737:chemistry*; D015800:Protozoan Proteins / Q000032:analysis*
D020543:Proteome; D015800:Protozoan Proteins