Logo Kérwá
 

Enzymatic labelling of venom phospholipase A2 toxins

Loading...
Thumbnail Image

Date

Authors

Spolaore, Barbara
Fernández Ulate, Julián
Lomonte, Bruno
Massimino, Maria Lina
Tonello, Fiorella

Journal Title

Journal ISSN

Volume Title

Publisher

Abstract

Almost all animal venoms contain secretory phospholipases A2 (PLA2s), 14 kDa disulfide-rich enzymes that hydrolyze membrane phospholipids at the sn-2 position, releasing lysophospholipids and fatty acids. These proteins, depending on their sequence, show a wide variety of biochemical, toxic and pharmacological effects and deserve to be studied for their numerous possible applications, and to improve antivenom drugs. The cellular localization and activity of a protein can be studied by conjugating it with a tag. In this work, we applied an enzymatic labelling method, using Streptomyces mobaraense transglutaminase, on three snake venom PLA2s: a recombinant neuro- and myotoxic group I PLA2 from Notechis scutatus scutatus, and two myotoxic group II PLA2s from Bothrops asper - one of them a natural catalytically inactive variant. We demonstrate that TGase can be used to produce active mono- or bi-derivatives of these three PLA2s modified at specific Lys residues, and that all three of these proteins, conjugated with fluorescent peptides, are internalized in primary myotubes.

Description

Keywords

Secretory phospholipase A2, Protein labelling, Enzymatic conjugation, Transglutaminase, snake venom

Citation

https://www.sciencedirect.com/science/article/pii/S0041010119304696?via%3Dihub

Collections

Endorsement

Review

Supplemented By

Referenced By