Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation
ABSTRACT Background Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect and the main vector of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). In the present study, the authors investigated whether a serine protease activity from the saliva of T. infestans has a role in vasomo...
Published in: | Journal of Venomous Animals and Toxins including Tropical Diseases |
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Online Access: | https://doi.org/10.1590/1678-9199-jvatitd-2020-0098 https://doaj.org/article/f65527f17c3a4f0fa0ff80027bbb178f |
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ftdoajarticles:oai:doaj.org/article:f65527f17c3a4f0fa0ff80027bbb178f 2023-05-15T15:15:16+02:00 Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation Karla A. Oliveira Ricardo J. S. Torquato Daniela C. G. Garcia Lustosa Tales Ribeiro Bruno W. L. Nascimento Lilian C. G. de Oliveira Maria A. Juliano Thaysa Paschoalin Virginia S. Lemos Ricardo N. Araujo Marcos H. Pereira Aparecida S. Tanaka 2021-03-01T00:00:00Z https://doi.org/10.1590/1678-9199-jvatitd-2020-0098 https://doaj.org/article/f65527f17c3a4f0fa0ff80027bbb178f EN eng SciELO http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992021000100304&tlng=en http://www.scielo.br/pdf/jvatitd/v27/1678-9199-jvatitd-27-e20200098.pdf https://doaj.org/toc/1678-9199 1678-9199 doi:10.1590/1678-9199-jvatitd-2020-0098 https://doaj.org/article/f65527f17c3a4f0fa0ff80027bbb178f Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 27 (2021) Triapsin Vasodilation PAR-2 receptor Nitric oxide Endothelial cells Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 article 2021 ftdoajarticles https://doi.org/10.1590/1678-9199-jvatitd-2020-0098 2022-12-31T11:04:32Z ABSTRACT Background Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect and the main vector of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). In the present study, the authors investigated whether a serine protease activity from the saliva of T. infestans has a role in vasomotor modulation, and in the insect-blood feeding by cleaving and activating protease-activated receptors (PARs). Methods T. infestans saliva was chromatographed as previously reported for purification of triapsin, a serine protease. The cleavage activity of triapsin on PAR peptides was investigated based on FRET technology. Mass spectrometry was used to analyze the sites of PAR-2 peptide cleaved by triapsin. NO measurements were performed using the DAN assay (2,3-diaminonapthalene). The vasorelaxant activity of triapsin was measured in vessels with or without functional endothelium pre-contracted with phenylephrine (3 µM). Intravital microscopy was used to assess the effect of triapsin on mouse skin microcirculation. Results Triapsin was able to induce hydrolysis of PAR peptides and showed a higher preference for cleavage of the PAR-2 peptide. Analysis by mass spectrometry confirmed a single cleavage site, which corresponds to the activation site of the PAR-2 receptor. Triapsin induced dose-dependent NO release in cultured human umbilical vein endothelial cells (HUVECs), reaching a maximum effect at 17.58 nM. Triapsin purified by gel-filtration chromatography (10-16 to 10-9 M) was applied cumulatively to mouse mesenteric artery rings and showed a potent endothelium-dependent vasodilator effect (EC30 = 10-12 M). Nitric oxide seems to be partially responsible for this vasodilator effect because L-NAME (L-NG-nitroarginine methyl ester 300 µM), a nitric oxide synthetase inhibitor, did not abrogate the vasodilation activated by triapsin. Anti-PAR-2 antibody completely inhibited vasodilation observed in the presence of triapsin activity. Triapsin activity also induced an increase in the mouse ear venular diameter. Conclusion ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Journal of Venomous Animals and Toxins including Tropical Diseases 27 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Triapsin Vasodilation PAR-2 receptor Nitric oxide Endothelial cells Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
spellingShingle |
Triapsin Vasodilation PAR-2 receptor Nitric oxide Endothelial cells Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 Karla A. Oliveira Ricardo J. S. Torquato Daniela C. G. Garcia Lustosa Tales Ribeiro Bruno W. L. Nascimento Lilian C. G. de Oliveira Maria A. Juliano Thaysa Paschoalin Virginia S. Lemos Ricardo N. Araujo Marcos H. Pereira Aparecida S. Tanaka Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation |
topic_facet |
Triapsin Vasodilation PAR-2 receptor Nitric oxide Endothelial cells Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
description |
ABSTRACT Background Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect and the main vector of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). In the present study, the authors investigated whether a serine protease activity from the saliva of T. infestans has a role in vasomotor modulation, and in the insect-blood feeding by cleaving and activating protease-activated receptors (PARs). Methods T. infestans saliva was chromatographed as previously reported for purification of triapsin, a serine protease. The cleavage activity of triapsin on PAR peptides was investigated based on FRET technology. Mass spectrometry was used to analyze the sites of PAR-2 peptide cleaved by triapsin. NO measurements were performed using the DAN assay (2,3-diaminonapthalene). The vasorelaxant activity of triapsin was measured in vessels with or without functional endothelium pre-contracted with phenylephrine (3 µM). Intravital microscopy was used to assess the effect of triapsin on mouse skin microcirculation. Results Triapsin was able to induce hydrolysis of PAR peptides and showed a higher preference for cleavage of the PAR-2 peptide. Analysis by mass spectrometry confirmed a single cleavage site, which corresponds to the activation site of the PAR-2 receptor. Triapsin induced dose-dependent NO release in cultured human umbilical vein endothelial cells (HUVECs), reaching a maximum effect at 17.58 nM. Triapsin purified by gel-filtration chromatography (10-16 to 10-9 M) was applied cumulatively to mouse mesenteric artery rings and showed a potent endothelium-dependent vasodilator effect (EC30 = 10-12 M). Nitric oxide seems to be partially responsible for this vasodilator effect because L-NAME (L-NG-nitroarginine methyl ester 300 µM), a nitric oxide synthetase inhibitor, did not abrogate the vasodilation activated by triapsin. Anti-PAR-2 antibody completely inhibited vasodilation observed in the presence of triapsin activity. Triapsin activity also induced an increase in the mouse ear venular diameter. Conclusion ... |
format |
Article in Journal/Newspaper |
author |
Karla A. Oliveira Ricardo J. S. Torquato Daniela C. G. Garcia Lustosa Tales Ribeiro Bruno W. L. Nascimento Lilian C. G. de Oliveira Maria A. Juliano Thaysa Paschoalin Virginia S. Lemos Ricardo N. Araujo Marcos H. Pereira Aparecida S. Tanaka |
author_facet |
Karla A. Oliveira Ricardo J. S. Torquato Daniela C. G. Garcia Lustosa Tales Ribeiro Bruno W. L. Nascimento Lilian C. G. de Oliveira Maria A. Juliano Thaysa Paschoalin Virginia S. Lemos Ricardo N. Araujo Marcos H. Pereira Aparecida S. Tanaka |
author_sort |
Karla A. Oliveira |
title |
Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation |
title_short |
Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation |
title_full |
Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation |
title_fullStr |
Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation |
title_full_unstemmed |
Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation |
title_sort |
proteolytic activity of triatoma infestans saliva associated with par-2 activation and vasodilation |
publisher |
SciELO |
publishDate |
2021 |
url |
https://doi.org/10.1590/1678-9199-jvatitd-2020-0098 https://doaj.org/article/f65527f17c3a4f0fa0ff80027bbb178f |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 27 (2021) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992021000100304&tlng=en http://www.scielo.br/pdf/jvatitd/v27/1678-9199-jvatitd-27-e20200098.pdf https://doaj.org/toc/1678-9199 1678-9199 doi:10.1590/1678-9199-jvatitd-2020-0098 https://doaj.org/article/f65527f17c3a4f0fa0ff80027bbb178f |
op_doi |
https://doi.org/10.1590/1678-9199-jvatitd-2020-0098 |
container_title |
Journal of Venomous Animals and Toxins including Tropical Diseases |
container_volume |
27 |
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