Barbara Spolaore - Academia.edu (original) (raw)
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Papers by Barbara Spolaore
Toxicon, 2019
Almost all animal venoms contain secretory phospholipases A 2 (PLA 2 s), 14 kDa disulfide-rich en... more Almost all animal venoms contain secretory phospholipases A 2 (PLA 2 s), 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 PLA 2 s: a recombinant neuro-and myotoxic group I PLA 2 from Notechis scutatus scutatus, and two myotoxic group II PLA 2 s 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 PLA 2 s modified at specific Lys residues, and that all three of these proteins, conjugated with fluorescent peptides, are internalized in primary myotubes.
Toxicon, 2019
Almost all animal venoms contain secretory phospholipases A 2 (PLA 2 s), 14 kDa disulfide-rich en... more Almost all animal venoms contain secretory phospholipases A 2 (PLA 2 s), 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 PLA 2 s: a recombinant neuro-and myotoxic group I PLA 2 from Notechis scutatus scutatus, and two myotoxic group II PLA 2 s 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 PLA 2 s modified at specific Lys residues, and that all three of these proteins, conjugated with fluorescent peptides, are internalized in primary myotubes.