Isabelle Florent, Fabien Lecaille, Jean-Jacques Montagne, Francis Gauthier, Joseph Schrével, Gilles Lalmanach
Labelling of four distinct trophozoite falcipains of Plasmodium falciparum by a cystatin-derived probe
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-CHN2) 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, falcipain-2? 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-2?. This activity-based probe represents a potential tool for profiling active falcipains in parasites.
Biological Chemistry, Walter de Gruyter
Print ISSN: 1431-6730
Volume: 386, 04/2005
Pages: 401 - 406
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