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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Specific calpain activity evaluation in Plasmodium parasites

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Gomes, Mayrim M. [1, 2] ; Budu, Alexandre [3] ; Ventura, Priscilla D. S. [2] ; Bagnaresi, Piero [3] ; Cotrin, Simone S. [3] ; Cunha, Rodrigo L. O. R. [4] ; Carmona, Adriana K. [3] ; Juliano, Luiz [3] ; Gazarini, Marcos L. [2]
Total Authors: 9
[1] Univ Fed Sao Paulo, Escola Paulista Med, Dept Microbiol Immunol & Parasitol, Sao Paulo - Brazil
[2] Univ Fed Sao Paulo, Dept Biociencias, Santos, SP - Brazil
[3] Univ Fed Sao Paulo, Dept Biofis, Sao Paulo - Brazil
[4] Univ Fed ABC, Ctr Ciencias Nat & Humanas, Santo Andre, SP - Brazil
Total Affiliations: 4
Document type: Journal article
Source: Analytical Biochemistry; v. 468, p. 22-27, JAN 1 2015.
Web of Science Citations: 5

In the intraerythrocytic trophozoite stages of Plasmodium falciparum, the calcium-dependent cysteine protease calpain (Pf-calpain) has an important role in the parasite calcium modulation and cell development. We established specific conditions to follow by confocal microscopy and spectrofluorimetry measurements the intracellular activity of Pf-calpain in live cells. The catalytic activity was measured using the fluorogenic Z-Phe-Arg-MCA (where Z is carbobenzoxy and MCA is 4-methylcoumaryl-7-amide). The calmodulin inhibitor calmidazolium and the sarcoplasmic reticulum calcium ATPase inhibitor thapsigargin were used for modifications in the cytosolic calcium concentrations that persisted in the absence of extracellular calcium. The observed calcium-dependent peptidase activity was greatly inhibited by specific cysteine protease inhibitor E-64 and by the selective calpain inhibitor ALLN (N-acetyl-t-leucyl-L-leucyl-L-norleucinal). Taken together, we observed that intracellular Pf-calpain can be selectively detected and is the main calcium-dependent protease in the intraerythrocytic stages of the parasite. The method described here can be helpful in cell metabolism studies and antimalarial drug screening. (C) 2014 Elsevier Inc. All rights reserved. (AU)

FAPESP's process: 13/12913-0 - Calcium-calmodulin of Plasmodium falciparum: identification of ligands and participation in the host-parasite signalling.
Grantee:Alexandre Budu
Support type: Scholarships in Brazil - Post-Doctorate
FAPESP's process: 11/15287-8 - Proteolytic activity study in Plasmodium and modulation of kallikrein-kinin system in infected livers
Grantee:Mayrim Machado Gomes Smaul
Support type: Scholarships in Brazil - Master
FAPESP's process: 10/09264-2 - Participation of phosphorylation and calcium homeostasis in intracellular proteolytic activity modulation in Plasmodium chabaudi
Grantee:Mayrim Machado Gomes Smaul
Support type: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 09/53840-0 - Fluorescence system for investigations of physiological and pathophysiological aspects in cellular models
Grantee:Adriana Karaoglanovic Carmona
Support type: Multi-user Equipment Program