Calpain-like: A Ca(2+) dependent cystein protease in Entamoeba histolytica cell death.

Abstract:

:Entamoeba histolytica programmed cell death (PCD) induced by G418 is characterized by the release of important amounts of intracellular calcium from reservoirs. Nevertheless, no typical caspases have been detected in the parasite, the PCD phenotype is inhibited by the cysteine protease inhibitor E-64. These results strongly suggest that Ca(2+)-dependent proteases could be involved in PCD. In this study, we evaluate the expression and activity of a specific dependent Ca(2+) protease, the calpain-like protease, by real-time quantitative PCR (RTq-PCR), Western blot assays and a enzymatic method during the induction of PCD by G418. Alternatively, using cell viability and TUNEL assays, we also demonstrated that the Z-Leu-Leu-Leu-al calpain inhibitor reduced the rate of cell death. The results demonstrated 4.9-fold overexpression of calpain-like gene 1.5 h after G418 PCD induction, while calpain-like protein increased almost two-fold with respect to basal calpain-like expression after 3 h of induction, and calpain activity was found to be approximately three-fold higher 6 h after treatment compared with untreated trophozoites. Taken together, these results suggest that this Ca(2+)-dependent protease could be involved in the executory phase of PCD.

journal_name

Exp Parasitol

authors

Monroy VS,Flores OM,García CG,Maya YC,Fernández TD,Pérez Ishiwara DG

doi

10.1016/j.exppara.2015.10.005

subject

Has Abstract

pub_date

2015-12-01 00:00:00

pages

245-51

eissn

0014-4894

issn

1090-2449

pii

S0014-4894(15)30052-7

journal_volume

159

pub_type

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