Expression of the First Recombinant Anti-Tumoral Snake Venom Kunitz-Type Serine Protease Inhibitor.
Maram MorjenWassim MoslahImen Touihri-BaraketiNajet Srairi-AbidJosé LuisNaziha MarrakchiJed JebaliPublished in: Toxins (2022)
PIVL is a Kunitz-type serine protease inhibitor that was previously characterized from Tunisian snake venom, Macrovipera lebetina transmediterranea . It reduced glioblastoma cells' development and significantly blocked angiogenesis in in-vitro and ex-vivo models. PIVL exerted these effects by interfering with αvβ3 integrin. In order to produce a biological active recombinant, the cDNA cloning and expression of PIVL was performed in Escherichia coli (BL21)-DE3 cells using pET-22b (+) vector. The recombinant PIVL protein (rPIVL) was purified by nickel affinity chromatography and has recognized monoclonal anti-His antibody. Functionally, rPIVL exhibited potent anti-tumor cell effects as well as anti-angiogenesis properties. Interestingly, we found that both native PIVL (nPIVL) and rPIVL modulated PI3/AKT and MAPK signaling pathways. In all, our results showed that we have successfully expressed the first active anti-oncogenic snake venom Kunitz-type protease inhibitor that can be a potential therapeutic drug against glioblastoma, in its native or recombinant form.
Keyphrases
- induced apoptosis
- signaling pathway
- escherichia coli
- poor prognosis
- cell cycle arrest
- cell free
- endothelial cells
- pi k akt
- oxidative stress
- mass spectrometry
- endoplasmic reticulum stress
- computed tomography
- single cell
- emergency department
- pet ct
- liquid chromatography
- multiple myeloma
- high speed
- bone marrow
- high performance liquid chromatography
- high resolution
- electronic health record
- cell adhesion
- amino acid
- carbon nanotubes
- solid phase extraction
- cell migration