Academic Journal
Specific calpain activity evaluation in Plasmodium parasites
| العنوان: | Specific calpain activity evaluation in Plasmodium parasites |
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| المؤلفون: | Gomes, Mayrim Machado UNIFESP, Budu, Alexandre UNIFESP, Ventura, Priscilla Dantas de Souza UNIFESP, Bagnaresi, Piero UNIFESP, Cotrin, Simone Silva UNIFESP, Cunha, Rodrigo Luiz Oliveira Rodrigues, Carmona, Adriana Karaoglanovic UNIFESP, Juliano, Luiz UNIFESP, Gazarini, Marcos Leoni UNIFESP |
| المساهمون: | Universidade Federal de São Paulo (UNIFESP), Universidade Federal do ABC (UFABC) |
| بيانات النشر: | Elsevier B.V. |
| سنة النشر: | 2015 |
| المجموعة: | Universidade Federal de São Paulo (UNIFESP): Repositório Institucional |
| مصطلحات موضوعية: | Plasmodium, Proteases, Calpain, Calcium signaling, Calmodulin |
| الوصف: | 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. ; Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) ; Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) ; Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Immunol & Parasitol, São Paulo, Brazil ; Universidade Federal de São Paulo, Dept Biofis, São Paulo, Brazil ; Universidade Federal de São Paulo, Dept Biociencias, Santos, SP, Brazil ; Univ Fed ABC, Ctr Ciencias Nat & Humanas, Santo Andre, SP, Brazil ; Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Immunol & Parasitol, São Paulo, Brazil ; Universidade Federal de São Paulo, Dept Biofis, São Paulo, Brazil ; Universidade Federal de São Paulo, Dept Biociencias, Santos, SP, Brazil ; FAPESP: 09/54598-9 ; FAPESP: 09/53840-0 ; FAPESP: ... |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | 22-27 |
| اللغة: | English |
| تدمد: | 0003-2697 |
| Relation: | Analytical Biochemistry; https://dx.doi.org/10.1016/j.ab.2014.09.005; Analytical Biochemistry. San Diego: Academic Press Inc Elsevier Science, v. 468, p. 22-27, 2015.; https://repositorio.unifesp.br/handle/11600/38577; WOS:000345811800004 |
| DOI: | 10.1016/j.ab.2014.09.005 |
| الاتاحة: | https://repositorio.unifesp.br/handle/11600/38577 https://doi.org/10.1016/j.ab.2014.09.005 |
| Rights: | Acesso restrito ; http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy |
| رقم الانضمام: | edsbas.584A5F0D |
| قاعدة البيانات: | BASE |
| تدمد: | 00032697 |
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| DOI: | 10.1016/j.ab.2014.09.005 |