Gelatin Zymography of Major Proteases in Exosomes of Leishmania major Promastigotes

Akram Azambakhtiar; Sedigheh Nabian; Mehdi  Mohebali; Mohammad   Taheri; Ramin Mazaheri  Nezhad Fard

doi:10.18502/ijpa.v18i4.14263

Akram Azambakhtiar Department of Parasitology, School of Veterinary Medicine, University of Tehran, Tehran, Iran
Sedigheh Nabian Department of Parasitology, School of Veterinary Medicine, University of Tehran, Tehran, Iran
Mehdi Mohebali Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Mohammad Taheri Rastegar Reference Laboratory, School of Veterinary Medicine, University of Tehran, Tehran, Iran
Ramin Mazaheri Nezhad Fard Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

DOI: https://doi.org/10.18502/ijpa.v18i4.14263

Keywords: Gelatin Zymography; Exosomes; Leishmania spp.; Iran

Abstract

Background: Enzymatic digestion of extra cellular matrix proteins by proteinases of Leishmania promastigotes is a complex process. Hence, studies on functional proteomics of these enzymes can help select these enzymes as possible vaccine candidates or selecting candidates for chemotherapy and immunotherapy. Several proteolytic enzymes are involved in virulence of Leishmania spp. These enzymes are mostly serine, cysteine and metalloproteases. We aimed to detect proteases in Leishmania promastigote exosomes.

Methods: Serine, cysteine and metalloproteases were investigated in exosomes and lysate of L. major promastigote using gelatin zymography. The study was carried out in the Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran, in 2021.

Results: Zymography findings of metalloproteinases showed transparent bands, including a 63-kDa glycoprotein (GP63). This glycoprotein is a major surface metalloproteinase. In addition, transparent bands belonged to serin proteases and cathepsin were demonstrated in gels associated to Leishmania promastigote lysate and exosomes.

Conclusion: Several metalloproteases, serin proteases and cathepsins were shown in promastigote lysate and exosomes of L. major, which could purified and used as fractions for immunodiagnostic.