JVATiTD - Articles

Official publication of CEVAP/UNESP
Submit your manuscript
Research

Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation

Karla A. Oliveira1, Ricardo J. S. Torquato2, Daniela C. G. Garcia Lustosa3, Tales Ribeiro4, Bruno W. L. Nascimento4, Lilian C. G. de Oliveira5, Maria A. Juliano5, Thaysa Paschoalin5, Virginia S. Lemos6, Ricardo N. Araujo4,7, Marcos H. Pereira4,7, Aparecida S. Tanaka2,7 [ + show more ]

J Venom Anim Toxins incl Trop Dis, 2021, 27:e20200098
Received: 26 June 2020 | Accepted: 28 January 2021 | Published online: 08 March 2021
https://doi.org/10.1590/1678-9199-JVATITD-2020-0098

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: Data from this study suggest a plausible association between triapsin activity mediated PAR-2 activation and vasodilation caused by T. infestans saliva.

 

Keywords: Triapsin; Vasodilation; PAR-2 receptor; Nitric oxide; Endothelial cells.

Full Article PDF
Top