Expression, purification and virucidal activity of two recombinant isoforms of phospholipase A2 from Crotalus durissus terrificus venom

Archives of Virology
Raquel Rinaldi RussoVictor Hugo Aquino

Abstract

The global emergence and re-emergence of arthropod-borne viruses (arboviruses) over the past four decades have become a public health crisis of international concern, especially in tropical and subtropical countries. A limited number of vaccines against arboviruses are available for use in humans; therefore, there is an urgent need to develop antiviral compounds. Snake venoms are rich sources of bioactive compounds with potential for antiviral prospection. The major component of Crotalus durissus terrificus venom is a heterodimeric complex called crotoxin, which is constituted by an inactive peptide (crotapotin) and a phospholipase A2 (PLA2-CB). We showed previously the antiviral effect of PLA2-CB against dengue virus, yellow fever virus and other enveloped viruses. The aims of this study were to express two PLA2-CB isoforms in a prokaryotic system and to evaluate their virucidal effects. The sequences encoding the PLA2-CB isoforms were optimized and cloned into a plasmid vector (pG21a) for recombinant protein expression. The recombinant proteins were expressed in the E. coli BL21(DE3) strain as insoluble inclusion bodies; therefore, the purification was performed under denaturing conditions, using urea for protein solubilizati...Continue Reading

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Citations

Jul 26, 2019·Current Topics in Medicinal Chemistry·Jéssica Peixoto RodriguesLuiz Ricardo Goulart
Sep 5, 2019·Pharmaceuticals·Juan-Carlos Saiz
Sep 11, 2020·Microorganisms·Rana Abdelnabi, Leen Delang
Jan 2, 2021·Frontiers in Microbiology·Débora Moraes de OliveiraAna Carolina Gomes Jardim
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Nov 17, 2020·Pathogens and Global Health·Caroline MwalikoZhiming Yuan
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Methods Mentioned

BETA
electrophoresis
blood collection
Protein Assay
Assay
gel filtration

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