A Plasmodium falciparum aminopeptidase gene belonging to the M1 family of zinc-metallopeptidases is expressed in erythrocytic stages.
A new single copy gene has been isolated from Plasmodium falciparum, by immunoscreening a genomic DNA expression library. The gene appears devoid of introns, displays the classical A + T richness and codon usage of P. falciparum genes, and is transcribed into a 4 kb mRNA in erythrocytic stages. The deduced amino acid sequence corresponds to a 1056 residue protein (122 kDa) containing the canonical HExxHx18E signature of zinc-metallopeptidase active sites of the M1 family at position 467-490, a downstream conserved tyrosine residue involved in catalysis in position 551, and the GAMEN conserved motif characteristic of aminopeptidases in the M1 family, at position 431-435. The greatest similarities were found with aminopeptidases N of Escherichia coli and Haemophilius influenza (more than 80% identical residues in the canonical signature of the active site) but significant similarities centred on the active site region exist with all other members of the M1 family such as other prokaryotic aminopeptidases, eukaryotic aminopeptidases A and N and leukotriene A4 hydrolases (40-50% identical residues in the canonical signature of the active site). A polyclonal serum raised to a synthetic peptide deduced from the gene labelled schizont proteins of 96 and 68 kDa purified to homogeneity and both displaying aminopeptidase activity, as well as cytoplasmic structures in schizont stages.
Florent
I
I
Laboratoire de Biologie Parasitaire, EP 1790 Biologie et Evolution des Parasites CNRS, Muséum National d'Histoire Naturelle, Paris, France. florent@mnhn.fr
Derhy
Z
Z
Allary
M
M
Monsigny
M
M
Mayer
R
R
Schrével
J
J
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004912:Erythrocytes / Q000469:parasitology*; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D015723:Gene Library; D017125:Genes, Protozoan; D008018:Life Cycle Stages / Q000502:physiology; D008666:Metalloendopeptidases / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment
D000626:Aminopeptidases; D008666:Metalloendopeptidases