Aminopeptidase N1 (EtAPN1), an M1 metalloprotease of the apicomplexan parasite Eimeria tenella, participates in parasite development.
Aminopeptidases N are metalloproteases of the M1 family that have been reported in numerous apicomplexan parasites, including Plasmodium, Toxoplasma, Cryptosporidium, and Eimeria. While investigating the potency of aminopeptidases as therapeutic targets against coccidiosis, one of the most important avian diseases caused by the genus Eimeria, we identified and characterized Eimeria tenella aminopeptidase N1 (EtAPN1). Its inhibition by bestatin and amastatin, as well as its reactivation by divalent ions, is typical of zinc-dependent metalloproteases. EtAPN1 shared a similar sequence, three-dimensional structure, and substrate specificity and similar kinetic parameters with A-M1 from Plasmodium falciparum (PfA-M1), a validated target in the treatment of malaria. EtAPN1 is synthesized as a 120-kDa precursor and cleaved into 96-, 68-, and 38-kDa forms during sporulation. Further, immunolocalization assays revealed that, similar to PfA-M1, EtAPN1 is present during the intracellular life cycle stages in both the parasite cytoplasm and the parasite nucleus. The present results support the hypothesis of a conserved role between the two aminopeptidases, and we suggest that EtAPN1 might be a valuable target for anticoccidiosis drugs.
INRA, UMR 1282, Infectiologie et Santé Publique, Nouzilly, France Université François Rabelais de Tours, UMR 1282, Infectiologie et Santé Publique, Tours, France.
Simon
S
Gras
Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wroclaw, Poland.
Anna
A
Byzia
INRA, UMR 1282, Infectiologie et Santé Publique, Nouzilly, France Université François Rabelais de Tours, UMR 1282, Infectiologie et Santé Publique, Tours, France.
Florence B
FB
Gilbert
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
Sheena
S
McGowan
Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wroclaw, Poland.
Marcin
M
Drag
INRA, UMR 1282, Infectiologie et Santé Publique, Nouzilly, France Université François Rabelais de Tours, UMR 1282, Infectiologie et Santé Publique, Tours, France anne.silvestre@tours.inra.fr.
Anne
A
Silvestre
INRA, UMR 1282, Infectiologie et Santé Publique, Nouzilly, France Université François Rabelais de Tours, UMR 1282, Infectiologie et Santé Publique, Tours, France.
Alisson
A
Niepceron
Université François Rabelais de Tours, INSERM, UMR 1100, Equipe Mécanismes Protéolytiques dans l'Inflammation, Centre d'Etude des Pathologies Respiratoires (CEPR), Tours, France.
Fabien
F
Lecaille
Université François Rabelais de Tours, INSERM, UMR 1100, Equipe Mécanismes Protéolytiques dans l'Inflammation, Centre d'Etude des Pathologies Respiratoires (CEPR), Tours, France.
Gilles
G
Lalmanach
INRA, UMR 1282, Infectiologie et Santé Publique, Nouzilly, France Université François Rabelais de Tours, UMR 1282, Infectiologie et Santé Publique, Tours, France.
Fabien
F
Brossier
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000981:Antiprotozoal Agents / Q000494:pharmacology; D016786:Eimeria tenella / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D045726:Metalloproteases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000494:pharmacology; D010802:Phylogeny; D011498:Protein Precursors / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D033761:Spores, Protozoan / Q000254:growth & development / Q000378:metabolism; D013379:Substrate Specificity
D000981:Antiprotozoal Agents; D010455:Peptides; D011498:Protein Precursors; D015800:Protozoan Proteins; C016977:amastatin; D045726:Metalloproteases; D000626:Aminopeptidases; D007930:Leucine; C012211:ubenimex