title: Inactivation of phospholipase A2 and metalloproteinase from Crotalus atrox venom by direct current.
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 0 and 8 V) into a reaction mixture for a few seconds. Then, we assayed the residual activity of phospholipases A(2) (PLA(2)),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 A(2) and metalloproteinases were consistently and irreversibly inactivated by direct current (between 0 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 A(2), homologue to snake venom phospholipases A(2), was also inactivated by direct current treatment. In order to understand the structural reasoning behind PLA(2) inactivation, circular dichroism measurements were conducted on homogeneous commercial pancreatic phospholipase A(2), and it was found that the enzyme undergoes structural alterations upon direct current exposure.
authors: Department of Biology, University of Genoa, V. le Bendetto XV, 3, 16132 Genova, Italy. Isabella.Panfoli@unige.it
authors: Isabella
authors: I
authors: Panfoli
authors: Silvia
authors: S
authors: Ravera
authors: Daniela
authors: D
authors: Calzia
authors: Elisa
authors: E
authors: Dazzi
authors: Simona
authors: S
authors: Gandolfo
authors: Isidoro M
authors: IM
authors: Pepe
authors: Alessandro
authors: A
authors: Morelli
mesh_terms: D000818:Animals; D002942:Circular Dichroism; D003435:Crotalid Venoms / Q000201:enzymology*; D017839:Crotalus; D004560:Electricity; D004789:Enzyme Activation; D054513:Group IV Phospholipases A2 / Q000378:metabolism*; D045726:Metalloproteases / Q000378:metabolism*; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D013552:Swine
chemical_list: D003435:Crotalid Venoms; D054513:Group IV Phospholipases A2; D010727:Phosphoric Diester Hydrolases; D045726:Metalloproteases