Characterization and cloning of the gene encoding the vacuolar membrane protein EXP-2 from Plasmodium falciparum.
As a contribution to the characterization of the parasitophorous vacuolar membrane from Plasmodium falciparum we have begun the identification of vacuolar membrane proteins. Exported protein-2 (EXP-2) is a vacuolar membrane protein exposed into the vacuolar space. To further characterize EXP-2, it was purified, and the 45 N-terminal amino acids were determined by micro-sequencing. Based on this information, partial cDNA and genomic fragments were amplified by PCR and used as probes for the isolation of complete cDNA and genomic DNA clones. The single copy gene is located on chromosome 14, and is transcribed during the ring stage of parasite development. The open reading frame encodes an N-terminal signal sequence which is cleaved from the mature protein. The amino acid composition of EXP-2 is characterized by charged amino acids, with a high abundance of aspartate residues in the C-terminal portion of the protein. In contrast to EXP-1, an integral protein of the vacuolar membrane, EXP-2 lacks a typical hydrophobic transmembrane domain. We suggest that EXP-2 may associate with the vacuolar membrane via an amphipathic helix located in the N-terminal half of the protein.
Fischer
K
K
Zentrum für Infektionsforschung, Universität Würzburg, Germany.
Marti
T
T
Rick
B
B
Johnson
D
D
Benting
J
J
Baumeister
S
S
Helmbrecht
C
C
Lanzer
M
M
Lingelbach
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D018076:DNA, Complementary; D015870:Gene Expression; D017125:Genes, Protozoan; D018503:Genome, Protozoan; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000235:genetics*; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics*; D017421:Sequence Analysis; D014617:Vacuoles / Q000235:genetics*
D018076:DNA, Complementary; C112548:EXP-2 protein, Plasmodium falciparum; D008565:Membrane Proteins; D015800:Protozoan Proteins