Cargando…

Cathepsin B-like and cell death in the unicellular human pathogen Leishmania

In several studies reporting cell death (CD) in lower eukaryotes and in the human protozoan parasite Leishmania, proteolytic activity was revealed using pan-caspase substrates or inhibitors such as carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK). However, most of the low...

Descripción completa

Detalles Bibliográficos
Autores principales: El-Fadili, A K, Zangger, H, Desponds, C, Gonzalez, I J, Zalila, H, Schaff, C, Ives, A, Masina, S, Mottram, J C, Fasel, N
Formato: Texto
Lenguaje:English
Publicado: Nature Publishing Group 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3032344/
https://www.ncbi.nlm.nih.gov/pubmed/21364675
http://dx.doi.org/10.1038/cddis.2010.51
_version_ 1782197452775882752
author El-Fadili, A K
Zangger, H
Desponds, C
Gonzalez, I J
Zalila, H
Schaff, C
Ives, A
Masina, S
Mottram, J C
Fasel, N
author_facet El-Fadili, A K
Zangger, H
Desponds, C
Gonzalez, I J
Zalila, H
Schaff, C
Ives, A
Masina, S
Mottram, J C
Fasel, N
author_sort El-Fadili, A K
collection PubMed
description In several studies reporting cell death (CD) in lower eukaryotes and in the human protozoan parasite Leishmania, proteolytic activity was revealed using pan-caspase substrates or inhibitors such as carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK). However, most of the lower eukaryotes do not encode caspase(s) but MCA, which differs from caspase(s) in its substrate specificity and cannot be accountable for the recognition of Z-VAD-FMK. In the present study, we were interested in identifying which enzyme was capturing the Z-VAD substrate. We show that heat shock (HS) induces Leishmania CD and leads to the intracellular binding of Z-VAD-FMK. We excluded binding and inhibition of Z-VAD-FMK to Leishmania major metacaspase (LmjMCA), and identified cysteine proteinase C (LmjCPC), a cathepsin B-like (CPC) enzyme, as the Z-VAD-FMK binding enzyme. We confirmed the specific interaction of Z-VAD-FMK with CPC by showing that Z-VAD binding is absent in a Leishmania mexicana strain in which the cpc gene was deleted. We also show that parasites exposed to various stress conditions release CPC into a soluble fraction. Finally, we confirmed the role of CPC in Leishmania CD by showing that, when exposed to the oxidizing agent hydrogen peroxide (H(2)O(2)), cpc knockout parasites survived better than wild-type parasites (WT). In conclusion, this study identified CPC as the substrate of Z-VAD-FMK in Leishmania and as a potential additional executioner protease in the CD cascade of Leishmania and possibly in other lower eukaryotes.
format Text
id pubmed-3032344
institution National Center for Biotechnology Information
language English
publishDate 2010
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-30323442011-02-24 Cathepsin B-like and cell death in the unicellular human pathogen Leishmania El-Fadili, A K Zangger, H Desponds, C Gonzalez, I J Zalila, H Schaff, C Ives, A Masina, S Mottram, J C Fasel, N Cell Death Dis Original Article In several studies reporting cell death (CD) in lower eukaryotes and in the human protozoan parasite Leishmania, proteolytic activity was revealed using pan-caspase substrates or inhibitors such as carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK). However, most of the lower eukaryotes do not encode caspase(s) but MCA, which differs from caspase(s) in its substrate specificity and cannot be accountable for the recognition of Z-VAD-FMK. In the present study, we were interested in identifying which enzyme was capturing the Z-VAD substrate. We show that heat shock (HS) induces Leishmania CD and leads to the intracellular binding of Z-VAD-FMK. We excluded binding and inhibition of Z-VAD-FMK to Leishmania major metacaspase (LmjMCA), and identified cysteine proteinase C (LmjCPC), a cathepsin B-like (CPC) enzyme, as the Z-VAD-FMK binding enzyme. We confirmed the specific interaction of Z-VAD-FMK with CPC by showing that Z-VAD binding is absent in a Leishmania mexicana strain in which the cpc gene was deleted. We also show that parasites exposed to various stress conditions release CPC into a soluble fraction. Finally, we confirmed the role of CPC in Leishmania CD by showing that, when exposed to the oxidizing agent hydrogen peroxide (H(2)O(2)), cpc knockout parasites survived better than wild-type parasites (WT). In conclusion, this study identified CPC as the substrate of Z-VAD-FMK in Leishmania and as a potential additional executioner protease in the CD cascade of Leishmania and possibly in other lower eukaryotes. Nature Publishing Group 2010-09 2010-09-02 /pmc/articles/PMC3032344/ /pubmed/21364675 http://dx.doi.org/10.1038/cddis.2010.51 Text en Copyright © 2010 Macmillan Publishers Limited http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under the Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Original Article
El-Fadili, A K
Zangger, H
Desponds, C
Gonzalez, I J
Zalila, H
Schaff, C
Ives, A
Masina, S
Mottram, J C
Fasel, N
Cathepsin B-like and cell death in the unicellular human pathogen Leishmania
title Cathepsin B-like and cell death in the unicellular human pathogen Leishmania
title_full Cathepsin B-like and cell death in the unicellular human pathogen Leishmania
title_fullStr Cathepsin B-like and cell death in the unicellular human pathogen Leishmania
title_full_unstemmed Cathepsin B-like and cell death in the unicellular human pathogen Leishmania
title_short Cathepsin B-like and cell death in the unicellular human pathogen Leishmania
title_sort cathepsin b-like and cell death in the unicellular human pathogen leishmania
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3032344/
https://www.ncbi.nlm.nih.gov/pubmed/21364675
http://dx.doi.org/10.1038/cddis.2010.51
work_keys_str_mv AT elfadiliak cathepsinblikeandcelldeathintheunicellularhumanpathogenleishmania
AT zanggerh cathepsinblikeandcelldeathintheunicellularhumanpathogenleishmania
AT despondsc cathepsinblikeandcelldeathintheunicellularhumanpathogenleishmania
AT gonzalezij cathepsinblikeandcelldeathintheunicellularhumanpathogenleishmania
AT zalilah cathepsinblikeandcelldeathintheunicellularhumanpathogenleishmania
AT schaffc cathepsinblikeandcelldeathintheunicellularhumanpathogenleishmania
AT ivesa cathepsinblikeandcelldeathintheunicellularhumanpathogenleishmania
AT masinas cathepsinblikeandcelldeathintheunicellularhumanpathogenleishmania
AT mottramjc cathepsinblikeandcelldeathintheunicellularhumanpathogenleishmania
AT faseln cathepsinblikeandcelldeathintheunicellularhumanpathogenleishmania