Labelling of four distinct trophozoite falcipains of Plasmodium falciparum by a cystatin-derived probe

Isabelle Florent (1) , Fabien Lecaille (2) , Jean-Jacques Montagne (3) , Francis Gauthier (4) , Joseph Schrével (1) , Gilles Lalmanach (2)

1 MCAM - Molécules de Communication et Adaptation des Micro-organismes
2 U618
3 IJM (UMR_7592) - Institut Jacques Monod
4 PRPA - Pathologies Respiratoires : Protéolyse et Aérosolthérapie

Trophozoite cysteine protease (TCP) activity, isolated from Plasmodium falciparum soluble 100 000 g extracts, displayed native falcipain-1 kinetic parameters towards peptidyl substrates. The labelling of either isolated TCP or soluble 100 000 g extracts by a cystatin-derived probe (biotinyl-Leu-Val-Gly-CHN 2) revealed a single band of ca. 30 kDa by SDS-PAGE, which was resolved into four spots displaying isoelectric points (pI) from 4.7 to 5.3 after two-dimensional separation. The molecular mass and pI correspond to those of falcipain-3, falcipain-2, fal-cipain-29 and falcipain-1, respectively. The two central spots were identified by matrix-assisted laser desorption/ ionisation time-of-flight mass spectrometry as falcipain-2 and falcipain-29. This activity-based probe represents a potential tool for profiling active falcipains in parasites.

Mots clés: activity-based probe; affinity labelling; cathepsin; cystatin; 2-D electrophoresis; malaria

Domaines: Biochimie [q-bio.BM]; Biologie moléculaire; Biologie cellulaire; Systématique, phylogénie et taxonomie; Protistologie; Microbiologie et Parasitologie; Biodiversité

Isabelle Florent, Fabien Lecaille, Jean-Jacques Montagne, Francis Gauthier, Joseph Schrével, et al.. Labelling of four distinct trophozoite falcipains of Plasmodium falciparum by a cystatin-derived probe. Biological Chemistry, 2005, 386 (4), pp.401 - 406. ⟨10.1515/BC.2005.048⟩. ⟨mnhn-02904301⟩