Plasmodium falciparum possesses a classical glutaredoxin and a second, glutaredoxin-like protein with a PICOT homology domain.
The genes coding for two different proteins with homologies to glutaredoxins have been identified in the genome of the malarial parasite Plasmodium falciparum. Both genes were amplified from a gametocytic cDNA and overexpressed in Escherichia coli. The smaller protein (named PfGrx-1) with 12.4 kDa in size exhibits the typical glutaredoxin active site motif "CPYC," shows glutathione-dependent glutaredoxin activity in the beta-hydroxyethyl disulfide (HEDS) assay, and reduces Trypanosoma brucei ribonucleotide reductase. Glutathione:HEDS transhydrogenase activity (approximately 60 milliunits/mg of protein) was clearly detectable in trophozoite extracts from eight different P. falciparum strains and did not differ between chloroquine-resistant and -sensitive parasites. Five different antimalarial drugs at 100 microm did not significantly influence isolated PfGrx-1 activity. In contrast, the second protein (deduced mass 19.9 kDa) with homology to glutaredoxins (31% identity to Schizosaccharomyces pombe in a 140-amino acid overlap) was not active in the HEDS assay; however, its general dithiol reducing activity was demonstrated in the insulin assay in the presence of dithiothreitol. Interestingly, the sequence contains a PICOT (for protein kinase C-interacting cousin of thioredoxin) homology domain, which might suggest regulatory functions of the protein. We named this protein PfGLP-1, for P. falciparum 1-Cys-glutaredoxin-like protein-1. In contrast to glutaredoxins, PfGLP-1 could not be reduced by glutathione. This is the first report on glutaredoxin-like proteins in the family of Plasmodia.
Rahlfs
S
S
Interdisciplinary Research Center, Justus Liebig University, 35392 Giessen, Germany.
Fischer
M
M
Becker
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000737:chemistry; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000032:analysis; D054477:Glutaredoxins; D007328:Insulin / Q000378:metabolism; D008969:Molecular Sequence Data; D010088:Oxidoreductases; D010963:Plasmodium falciparum / Q000737:chemistry / Q000235:genetics*; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012264:Ribonucleotide Reductases / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D002352:Carrier Proteins; D016054:DNA, Protozoan; D054477:Glutaredoxins; C438386:Grx-1 protein, Plasmodium falciparum; D007328:Insulin; D011506:Proteins; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D010088:Oxidoreductases; D012264:Ribonucleotide Reductases