To achieve our aim of understanding the interactions between direct current and enzymes in solution, we exposed reconstituted Crotalus atrox venom to direct electric current by immersing two platinum thread electrodes connected to a voltage generator (between O and 8 V) into a reaction mixture for a few seconds. Then, we assayed the residual activity of phospholipases A2 (PLA2),metalloproteinases, and phosphodiesterases, abundant in crotaline snake venoms and relevant in the pathophysiology of envenomation, characterized by hemorrhage, pain, and tissue damage. C. atrox venom phospholipase Aa and metalloproteinases were consistently and irreversibly inactivated by direct current (between O and 0.7 mA) exposure. In contrast, C. atrox venom phosphodiesterases were not affected. Total protein content and temperature of the sample remained the same. Secretory pancreatic phospholipase A2, homologue to snake venom phospholipases A 2, was also inactivated by direct current treatment. In order to understand the structural reasoning behind PLAa inactivation, circular dichroism measurements were conducted on homogeneous commercial pancreatic phospholipase A2, and it was found that the enzyme undergoes structural alterations upon direct current exposure. © 2007 Wiley Periodicals, Inc.

Inactivation of phospholipase A2 and metalloproteinase from Crotalus atrox venom by direct current

PANFOLI, ISABELLA;RAVERA, SILVIA;CALZIA, DANIELA;PEPE, ISIDORO;MORELLI, ALESSANDRO
2007-01-01

Abstract

To achieve our aim of understanding the interactions between direct current and enzymes in solution, we exposed reconstituted Crotalus atrox venom to direct electric current by immersing two platinum thread electrodes connected to a voltage generator (between O and 8 V) into a reaction mixture for a few seconds. Then, we assayed the residual activity of phospholipases A2 (PLA2),metalloproteinases, and phosphodiesterases, abundant in crotaline snake venoms and relevant in the pathophysiology of envenomation, characterized by hemorrhage, pain, and tissue damage. C. atrox venom phospholipase Aa and metalloproteinases were consistently and irreversibly inactivated by direct current (between O and 0.7 mA) exposure. In contrast, C. atrox venom phosphodiesterases were not affected. Total protein content and temperature of the sample remained the same. Secretory pancreatic phospholipase A2, homologue to snake venom phospholipases A 2, was also inactivated by direct current treatment. In order to understand the structural reasoning behind PLAa inactivation, circular dichroism measurements were conducted on homogeneous commercial pancreatic phospholipase A2, and it was found that the enzyme undergoes structural alterations upon direct current exposure. © 2007 Wiley Periodicals, Inc.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11567/249135
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