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Arrested growth of Trypanosoma cruzi by the calpain inhibitor MDL28170 and detection of calpain homologues in epimastigote forms

  • L. S. SANGENITO (a1), V. ENNES-VIDAL (a2), F. A. MARINHO (a1), F. F. DA MOTA (a3), A. L. S. SANTOS (a1), C. M. D'AVILA-LEVY (a2) and M. H. BRANQUINHA (a1)...
Abstract
SUMMARY

In this paper, we aimed to explore the effects of the calpain inhibitor III (MDL28170) and to detect calpain-like molecules (CALPs) in epimastigote forms of Trypanosoma cruzi isolate Dm28c. MDL28170 at 70 μM promoted a powerful reduction in the growth rate after 48 h. The IC50 value was calculated to be 31·7 μM. This inhibitor promoted an increase in the cellular volume, but not cell lysis, resulting in a trypanostatic effect. T. cruzi CALPs presented a strong cross-reactivity with anti-Drosophila melanogaster calpain and anti-cytoskeleton-associated protein from Trypanosoma brucei antibodies, and labelling was found mainly intracellularly. Furthermore, an 80 kDa reactive protein was detected by Western blotting assays. No significant cross-reactivity was found with anti-human brain calpain antibody. The expression of CALPs was decreased in cells kept for long periods in axenic cultures in comparison to a strain recently isolated from mice, as well as in MDL28170-treated cells, the latter being paralleled by an increased expression of cruzipain. Different levels of CALPs expression were also detected in distinct phylogenetic lineages, like Y strain (lineage TCI), Dm28c (TCII) and INPA6147 strain (Z3 zymodeme). These results may contribute for the investigation of the functions of CALPs in trypanosomatids.

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Corresponding author
*Corresponding author: Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes, Universidade Federal do Rio de Janeiro (UFRJ), Centro de Ciências da Saúde (CCS), Av. Carlos Chagas Filho, 373, Bloco I, Cidade Universitária, Rio de Janeiro, RJ, 21941-902, Brazil. Tel: +55 21 2562 6743. Fax: +55 21 2560 8344. E-mail: mbranquinha@micro.ufrj.br
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H. M. Andrade , S. M. F. Murta , A. Chapeaurouge , J. Perales , P. Nirdé and A. Romanha (2008). Proteomic analysis of Trypanosoma cruzi resistance to benznidazole. Journal of Proteome Research 7, 23572367.

F. Battaglia , F. Trinchese , S. Liu , S. Walter , R. A. Nixon and O. Arancio (2003). Calpain inhibitors, a treatment for Alzheimer's disease: position paper. Journal of Molecular Neurosciences 20, 357362.

J. J. Cazzulo , V. Stoka and V. Turk (2001). The major cysteine proteinase of Trypanosoma cruzi: a valid target for chemotherapy of Chagas disease. Current Pharmacological Design 7, 11431156.

V. T. Contreras , A. R. Lima and G. Zorrilla (1998). Trypanosoma cruzi: maintenance in culture modify gene and antigenic expression of metacyclic trypomastigotes. Memórias do Instituto Oswaldo Cruz 93, 753760.

C. M. d'Avila-Levy , F. A. Marinho , L. O. Santos , J. L. M. Martins , A. L. S. Santos and M. H. Branquinha (2006 a). Antileishmanial activity of MDL28170, a potent calpain inhibitor. International Journal of Antimicrobial Agents 28, 138142.

C. M. d'Avila-Levy , F. A. Dias , A. C. Nogueira de Melo , J. L. Martins , A. H. C. S. Lopes , A. L. S. Santos , A. B. Vermelho and M. H. Branquinha (2006 b). Insights into the role of gp63-like proteins in lower trypanosomatids. FEMS Microbiology Letters 254, 149156.

C. M. d'Avila-Levy , R. F. Souza , R. C. Gomes , A. B. Vermelho and M. H. Branquinha (2003). A novel extracellular cysteine proteinase from Crithidia deanei. Archives of Biochemistry and Biophysics 420, 18.

K. Ersfeld , H. Barraclough and K. Gull (2005). Evolutionary relationships and protein domain architecture in an expanded calpain superfamily in kinetoplastid parasites. The Journal of Molecular Evolution 61, 742757.

V. Giese , B. Dallagiovanna , F. K. Marchini , D. P. Pavoni , M. A. Krieger and S. Goldenberg (2008). Trypanosoma cruzi: a stage-specific calpain-like protein is induced after various kinds of stress. Memórias do Instituto Oswaldo Cruz 103, 598601.

D. E. Goll , V. F. Thompson , H. Li , W. Wei and J. Cong (2003). The calpain system. Physiological Reviews 83, 731801.

F. Grynspan , W. R. Griffin , A. Cataldo , S. Katayama and R. A. Nixon (1997). Active site-directed antibodies identify calpain II as an early-appearing and pervasive component of neurofibrillary pathology in Alzheimer's disease. Brain Research 763, 145158.

C. Hertz-Fowler , K. Ersfeld and K. Gull (2001). CAP5.5, a life-cycle-regulated, cytoskeleton-associated protein is a member of a novel family of calpain-related proteins in Trypanosoma brucei. Molecular and Biochemical Parasitology 116, 2534.

R. R. Mendonça-Filho , I. A. Rodrigues , D. S. Alviano , A. L. S. Santos , R. M. A. Soares , C. S. Alviano , A. H. C. S. Lopes and M. S. Rosa (2004). Leishmanicidal activity of polyphenolic-rich extract from husk fiber of Cocos nucifera Linn. (Palmae). Research in Microbiology 155, 136143.

M. A. Miles , A. Souza , M. Povoa , J. J. Shaw , R. Lainson and P. J. Toye (1978). Isozymic heterogeneity of Trypanosoma cruzi in the first autochtonous patients with Chagas’ disease in Amazonian Brazil. Nature, London 272, 819821.

A. Rami , D. Ferger and J. Krieglstein (1997). Blockade of calpain proteolytic activity rescues neurons from glutamate excitotoxicity. Neuroscience Research 27, 9397.

C. S. Ramos , F. A. L. Franco , D. F. Smith and S. R. B. Uliana (2004). Characterisation of a new Leishmania META gene and genomic analysis of the META cluster. FEMS Microbiology Letters 238, 213219.

P. Salotra , R. C. Duncan , R. Singh , B. V. S. Raju , G. Sreenivas and H. L. Nakhasi (2006). Upregulation of surface proteins in Leishmania donovani isolated from patients of post kala-azar dermal leishmaniasis. Microbes and Infection 8, 637644.

A. L. S. Santos , C. M. d'Avila-Levy , F. A. Dias , R. O. Ribeiro , F. M. Pereira , C. G. Elias , T. Souto-Padrón , A. H. Lopes , C. S. Alviano , M. H. Branquinha and R. M. A. Soares (2006). Phytomonas serpens: cysteine peptidase inhibitors interfere with growth, ultrastructure and host adhesion. International Journal for Parasitology 36, 4756.

A. Saxena , E. A. Worthey , S. Yan , A. Leland , K. D. Stuart and P. J. Myler (2003). Evaluation of differential gene expression in Leishmania major Friedlin procyclics and metacyclics using DNA microarray analysis. Molecular and Biochemical Parasitology 129, 103114.

R. P. Souto , O. Fernandes , A. M. Macedo , D. A. Campbell and B. Zingales (1996). DNA markers define two major phylogenetic lineages of Trypanosoma cruzi. Molecular and Biochemical Parasitology 83, 141152.

A. M. Tomas , M. A. Miles and J. M. Kelly (1997). Overexpression of cruzipain, the major cysteine proteinase of Trypanosoma cruzi, is associated with enhanced metacyclogenesis. European Journal of Biochemistry 244, 596603.

L. Troeberg , R. E. Morty , R. N. Pike , J. D. Lonsdale-Eccles , J. T. Palmer , J. H. McKerrow and T. H. Coetzer (1999). Cysteine proteinase inhibitors kill cultured bloodstream forms of Trypanosoma brucei brucei. Experimental Parasitology 91, 349355.

B. Vergnes , B. Gourbal , I. Gorard , S. Sundar , J. Drummelsmith and M. Ouellette (2007). A proteomics screen implicates HSP83 and a small kinetoplastid calpain-related protein in drug resistance in Leishmania donovani clinical field isolates by modulating drug-induced programmed cell death. Molecular Cell Proteomics 6, 88101.

A. B. Vermelho , S. Giovanni-de-Simone , C. M. d'Avila-Levy , A. L. S. Santos , A. C. N. Melo , F. P. Silva Jr,, E. P. S. Bom and M. H. Branquinha (2007). Trypanosomatidae peptidases: a target for drugs development. Current Enzyme Inhibition 3, 1948.

V. Yong , V. Schmitz , M. A. Vannier-Santos , A. P. C. A. Lima , G. Lalmanach , L. Juliano , F. Gauthier and J. Scharfstein (2000). Altered expression of cruzipain and a cathepsin B-like target in a Trypanosoma cruzi cell line displaying resistance to synthetic inhibitors of cysteine-proteinases. Molecular and Biochemical Parasitology 109, 4759.

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Parasitology
  • ISSN: 0031-1820
  • EISSN: 1469-8161
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