Analysis of membrane proteins by two-dimensional electrophoresis: comparison of the proteins extracted from normal or Plasmodium falciparum-infected erythrocyte ghosts.
Parasite-encoded membrane proteins translocated to the surface of infected erythrocytes or in specialized vesicles underneath (Maurer's clefts) play a key role in the asexual life cycle of Plasmodium falciparum (a malaria-causing protozoan), by mediating key steps such as red blood cell invasion, sequestration of infected cells in microcapillaries, and red blood cell rupture. A large-scale analysis of these membrane proteins would therefore be of great help to gain knowledge of the different stages of the Plasmodium falciparum life cycle. In order to be able to detect and identify parasite-encoded proteins directed to the red blood cell membrane, we first defined the conditions required for optimal extraction and separation of normal red blood cell ghost proteins by two-dimensional gel electrophoresis. These conditions included the use of urea, thiourea and new zwitterionic detergents in the extraction and isoelectric focusing media. The optimized conditions were then applied to analyze normal and P. falciparum-infected red blood cell ghosts. Several protein spots were found only in infected ghosts and are expected to represent parasite-encoded proteins. These proteins are currently under investigation.
Rabilloud
T
T
CEA-Laboratoire de Bioénergétique Cellulaire et Pathologique, Grenoble, France. thierry@sanrafael.ceng.cea.fr
Blisnick
T
T
Heller
M
M
Luche
S
S
Aebersold
R
R
Lunardi
J
J
Braun-Breton
C
C
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004910:Erythrocyte Membrane / Q000737:chemistry* / Q000469:parasitology; D006801:Humans; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis*; D010963:Plasmodium falciparum / Q000302:isolation & purification*
D008565:Membrane Proteins