Use of a phosphopeptide as a ligand to purify phospholipase A2 from the venom of Crotalus durisuss terrificus by affinity chromatography

SAAVEDRA, SOLEDAD L.; GIUDICESSI, SILVANA L.; CASCONE, OSVALDO; ACOSTA, GERARDO; CAMPERI, SILVIA A.; MARTINEZ CERON, MARIA C.; AVILA, LUCÍA; ALBERICIO, FERNANDO

Journal of Chromatography B

ELSEVIER SCIENCE BV

Lugar: Amsterdam; Año: 2020 vol. 1146 p. 122070 - 122076

1570-0232

The venom of the South American snake Crotalus durissus terrificus (Cdt) is asource of a wide variety of toxins, some of them with interesting harmacologicalapplications. Among them, the phospholipase A2 (PLA2) subunit of crotoxin (Ctx) has been studied for its possible use as an antiviral, antibacterial and antitumoral agent. Peptides have proven to be very useful ligands for the purification of numerous molecules such as antibodies, toxins, enzymes and other proteins. Here, we aimed to use a phosphopeptide (PLys) as a ligand for PLA2 purification. The synthesis was carried out on solid phase on RinkAmide-ChemMatrix resin and then P-Lys was immobilized on NHS-agarose and evaluatedas a chromatographic matrix. Under the best conditions, the percentage of total protein adsorption reached 39% and only the eluate fraction presented PLA2 enzymatic activity. The analysis by SDS-PAGE of the eluate showed three bands, one corresponding to the molecular weight of PLA2 (14 kDa). Said bands were analyzed by mass spectrometry and identified as PLA2 and its oligomers. The purity was over 90%. In addition, the process also allowed the isolation of crotamine.