2. Academic Validation
  3. Structural, enzymatic and pharmacological profiles of AplTX-II - A basic sPLA2 (D49) isolated from the Agkistrodon piscivorus leucostoma snake venom

Structural, enzymatic and pharmacological profiles of AplTX-II - A basic sPLA2 (D49) isolated from the Agkistrodon piscivorus leucostoma snake venom

  • Int J Biol Macromol. 2021 Apr 1:175:572-585. doi: 10.1016/j.ijbiomac.2021.01.187.
Letícia M Resende 1,  José R Almeida 2,  Tatiana A Guaraca-Medina 2,  Matilde F Viegas 3,  Andreimar M Soares 4,  Maria J Ramos 3,  Pedro A Fernandes 3,  Sergio Marangoni 1,  Saulo L Da Silva 5
Affiliations

Affiliations

  • 1 Department of Biochemistry and Tissue Biology, Institute of Biology, Campinas State University (UNICAMP), Campinas, SP, Brazil.
  • 2 IKIAM - Universidad Regional Amazonica, Km 7 Via Muyuna, Tena, Napo, Ecuador.
  • 3 LAQV/Requimte, University of Porto, Rua do Campo Alegre s/n, Porto, Portugal; Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, Rua do Campo Alegre s/n, Porto, Portugal.
  • 4 Biotechnology Laboratory for Proteins and Bioactive Compounds from the Western Amazon, Oswaldo Cruz Foundation Rondonia, Porto Velho, Rondonia, Brazil; Oswaldo Cruz Foundation, FIOCRUZ, Rondonia and Federal University of Rondonia, UNIR, Porto Velho, RO, Brazil.
  • 5 LAQV/Requimte, University of Porto, Rua do Campo Alegre s/n, Porto, Portugal; Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, Rua do Campo Alegre s/n, Porto, Portugal; Faculty of Chemical Sciences, University of Cuenca, Cuenca, Azuay, Ecuador. Electronic address: saulo.da.silva@ucuenca.edu.ec.
PMID: 33529631 DOI: 10.1016/j.ijbiomac.2021.01.187
Abstract

A basic sPLA2 (D49) from the venom of snake Agkistrodon piscivorus leucostoma (AplTX-II) was isolated, purified and characterized. We determined the enzymatic and pharmacological profiles of this toxin. AplTX-II was isolated with a high level of purity through reverse phase chromatography and molecular exclusion. The Enzyme showed pI 9.48 and molecular weight of 14,003 Da. The enzymatic activity of the AplTX-II depended on CA2+ pH and temperature. The comparison of the primary structure with Other sPLA2s revealed that AplTX-II presented all the structural reasons expected for a basic sPLA2s. Additionally, we have resolved its structure with the docked synthetic substrate NOBA (4-nitro-3-octanoyloxy benzoic acid) by homology modeling, and performed MD simulations with explicit solvent. Structural similarities were found between the enzyme's modeled structure and Other snake sPLA2 X-Ray structures, available in the PDB database. NOBA and active-site water molecules spontaneously adopted stable positions and established interactions in full agreement with the reaction mechanism, proposed for the physiological substrate, suggesting that NOBA hydrolysis is an excellent model to study phospholipid hydrolysis.

Keywords

Agkistrodon piscivorus leuscostoma; Basic sPLA(2) D49; Molecular modeling; NOBA (4-nitro-3-octanoyloxy benzoic acid).

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