Cargando…

Crovirin, a Snake Venom Cysteine-Rich Secretory Protein (CRISP) with Promising Activity against Trypanosomes and Leishmania

BACKGROUND: The neglected human diseases caused by trypanosomatids are currently treated with toxic therapy with limited efficacy. In search for novel anti-trypanosomatid agents, we showed previously that the Crotalus viridis viridis (Cvv) snake venom was active against infective forms of Trypanosom...

Descripción completa

Detalles Bibliográficos
Autores principales: Adade, Camila M., Carvalho, Ana Lúcia O., Tomaz, Marcelo A., Costa, Tatiana F. R., Godinho, Joseane L., Melo, Paulo A., Lima, Ana Paula C. A., Rodrigues, Juliany C. F., Zingali, Russolina B., Souto-Padrón, Thaïs
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4199522/
https://www.ncbi.nlm.nih.gov/pubmed/25330220
http://dx.doi.org/10.1371/journal.pntd.0003252
_version_ 1782339922180440064
author Adade, Camila M.
Carvalho, Ana Lúcia O.
Tomaz, Marcelo A.
Costa, Tatiana F. R.
Godinho, Joseane L.
Melo, Paulo A.
Lima, Ana Paula C. A.
Rodrigues, Juliany C. F.
Zingali, Russolina B.
Souto-Padrón, Thaïs
author_facet Adade, Camila M.
Carvalho, Ana Lúcia O.
Tomaz, Marcelo A.
Costa, Tatiana F. R.
Godinho, Joseane L.
Melo, Paulo A.
Lima, Ana Paula C. A.
Rodrigues, Juliany C. F.
Zingali, Russolina B.
Souto-Padrón, Thaïs
author_sort Adade, Camila M.
collection PubMed
description BACKGROUND: The neglected human diseases caused by trypanosomatids are currently treated with toxic therapy with limited efficacy. In search for novel anti-trypanosomatid agents, we showed previously that the Crotalus viridis viridis (Cvv) snake venom was active against infective forms of Trypanosoma cruzi. Here, we describe the purification of crovirin, a cysteine-rich secretory protein (CRISP) from Cvv venom with promising activity against trypanosomes and Leishmania. METHODOLOGY/PRINCIPAL FINDINGS: Crude venom extract was loaded onto a reverse phase analytical (C8) column using a high performance liquid chromatographer. A linear gradient of water/acetonitrile with 0.1% trifluoroacetic acid was used. The peak containing the isolated protein (confirmed by SDS-PAGE and mass spectrometry) was collected and its protein content was measured. T. cruzi trypomastigotes and amastigotes, L. amazonensis promastigotes and amastigotes and T. brucei rhodesiense procyclic and bloodstream trypomastigotes were challenged with crovirin, whose toxicity was tested against LLC-MK(2) cells, peritoneal macrophages and isolated murine extensor digitorum longus muscle. We purified a single protein from Cvv venom corresponding, according to Nano-LC MS/MS sequencing, to a CRISP of 24,893.64 Da, henceforth referred to as crovirin. Human infective trypanosomatid forms, including intracellular amastigotes, were sensitive to crovirin, with low IC(50) or LD(50) values (1.10–2.38 µg/ml). A considerably higher concentration (20 µg/ml) of crovirin was required to elicit only limited toxicity on mammalian cells. CONCLUSIONS: This is the first report of CRISP anti-protozoal activity, and suggests that other members of this family might have potential as drugs or drug leads for the development of novel agents against trypanosomatid-borne neglected diseases.
format Online
Article
Text
id pubmed-4199522
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-41995222014-10-21 Crovirin, a Snake Venom Cysteine-Rich Secretory Protein (CRISP) with Promising Activity against Trypanosomes and Leishmania Adade, Camila M. Carvalho, Ana Lúcia O. Tomaz, Marcelo A. Costa, Tatiana F. R. Godinho, Joseane L. Melo, Paulo A. Lima, Ana Paula C. A. Rodrigues, Juliany C. F. Zingali, Russolina B. Souto-Padrón, Thaïs PLoS Negl Trop Dis Research Article BACKGROUND: The neglected human diseases caused by trypanosomatids are currently treated with toxic therapy with limited efficacy. In search for novel anti-trypanosomatid agents, we showed previously that the Crotalus viridis viridis (Cvv) snake venom was active against infective forms of Trypanosoma cruzi. Here, we describe the purification of crovirin, a cysteine-rich secretory protein (CRISP) from Cvv venom with promising activity against trypanosomes and Leishmania. METHODOLOGY/PRINCIPAL FINDINGS: Crude venom extract was loaded onto a reverse phase analytical (C8) column using a high performance liquid chromatographer. A linear gradient of water/acetonitrile with 0.1% trifluoroacetic acid was used. The peak containing the isolated protein (confirmed by SDS-PAGE and mass spectrometry) was collected and its protein content was measured. T. cruzi trypomastigotes and amastigotes, L. amazonensis promastigotes and amastigotes and T. brucei rhodesiense procyclic and bloodstream trypomastigotes were challenged with crovirin, whose toxicity was tested against LLC-MK(2) cells, peritoneal macrophages and isolated murine extensor digitorum longus muscle. We purified a single protein from Cvv venom corresponding, according to Nano-LC MS/MS sequencing, to a CRISP of 24,893.64 Da, henceforth referred to as crovirin. Human infective trypanosomatid forms, including intracellular amastigotes, were sensitive to crovirin, with low IC(50) or LD(50) values (1.10–2.38 µg/ml). A considerably higher concentration (20 µg/ml) of crovirin was required to elicit only limited toxicity on mammalian cells. CONCLUSIONS: This is the first report of CRISP anti-protozoal activity, and suggests that other members of this family might have potential as drugs or drug leads for the development of novel agents against trypanosomatid-borne neglected diseases. Public Library of Science 2014-10-16 /pmc/articles/PMC4199522/ /pubmed/25330220 http://dx.doi.org/10.1371/journal.pntd.0003252 Text en © 2014 Adade et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Adade, Camila M.
Carvalho, Ana Lúcia O.
Tomaz, Marcelo A.
Costa, Tatiana F. R.
Godinho, Joseane L.
Melo, Paulo A.
Lima, Ana Paula C. A.
Rodrigues, Juliany C. F.
Zingali, Russolina B.
Souto-Padrón, Thaïs
Crovirin, a Snake Venom Cysteine-Rich Secretory Protein (CRISP) with Promising Activity against Trypanosomes and Leishmania
title Crovirin, a Snake Venom Cysteine-Rich Secretory Protein (CRISP) with Promising Activity against Trypanosomes and Leishmania
title_full Crovirin, a Snake Venom Cysteine-Rich Secretory Protein (CRISP) with Promising Activity against Trypanosomes and Leishmania
title_fullStr Crovirin, a Snake Venom Cysteine-Rich Secretory Protein (CRISP) with Promising Activity against Trypanosomes and Leishmania
title_full_unstemmed Crovirin, a Snake Venom Cysteine-Rich Secretory Protein (CRISP) with Promising Activity against Trypanosomes and Leishmania
title_short Crovirin, a Snake Venom Cysteine-Rich Secretory Protein (CRISP) with Promising Activity against Trypanosomes and Leishmania
title_sort crovirin, a snake venom cysteine-rich secretory protein (crisp) with promising activity against trypanosomes and leishmania
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4199522/
https://www.ncbi.nlm.nih.gov/pubmed/25330220
http://dx.doi.org/10.1371/journal.pntd.0003252
work_keys_str_mv AT adadecamilam crovirinasnakevenomcysteinerichsecretoryproteincrispwithpromisingactivityagainsttrypanosomesandleishmania
AT carvalhoanaluciao crovirinasnakevenomcysteinerichsecretoryproteincrispwithpromisingactivityagainsttrypanosomesandleishmania
AT tomazmarceloa crovirinasnakevenomcysteinerichsecretoryproteincrispwithpromisingactivityagainsttrypanosomesandleishmania
AT costatatianafr crovirinasnakevenomcysteinerichsecretoryproteincrispwithpromisingactivityagainsttrypanosomesandleishmania
AT godinhojoseanel crovirinasnakevenomcysteinerichsecretoryproteincrispwithpromisingactivityagainsttrypanosomesandleishmania
AT melopauloa crovirinasnakevenomcysteinerichsecretoryproteincrispwithpromisingactivityagainsttrypanosomesandleishmania
AT limaanapaulaca crovirinasnakevenomcysteinerichsecretoryproteincrispwithpromisingactivityagainsttrypanosomesandleishmania
AT rodriguesjulianycf crovirinasnakevenomcysteinerichsecretoryproteincrispwithpromisingactivityagainsttrypanosomesandleishmania
AT zingalirussolinab crovirinasnakevenomcysteinerichsecretoryproteincrispwithpromisingactivityagainsttrypanosomesandleishmania
AT soutopadronthais crovirinasnakevenomcysteinerichsecretoryproteincrispwithpromisingactivityagainsttrypanosomesandleishmania