title: Coupling reaction and properties of poly(ethylene glycol)-linked phospholipases A2.
abstract: Secretory phospholipases A2 (PLA2) from Naja naja naja (cobra snake) venom, from Bothrops neuwiedii (crotalid snake) venom (two isoforms) and from bee venom were modified with tresylated monomethoxy poly(ethylene glycol) (TMPEG). The kinetic and inflammatory properties of the adducts (PEG-PLA2) were measured. As found by gel permeation chromatography, 95-100% of P-1 PLA2 from B. neuwiedii and PLA2 from N. naja naja venom change their chromatographic mobility after TMPEG treatment. By contrast, only 50-60% of both P-3-PLA2 from B. neuwiedii and PLA2 from bee venom modify their elution profile from Superdex 75. All the modified proteins preserved the enzymatic activity toward phospholipid monolayers, but with a reduced specific activity and greater lag times than the unmodified controls. These results suggest that the PEG-PLA2 complexes would have an altered interaction with lipid membranes. The PEG-linked proteins preserve their edema-inducing activity evaluated by the rat hind-paw edema test except for N. naja naja PEG-PLA2 in which inflammatory activity was significatively decreased. Altogether, the results show a partial dissociation of catalytic and inflammatory activities of Group II and III secretory PLA2s after their modification with PEG.
authors.affiliation: CEPROCOR-Agencia Córdoba Ciencia S.E., Argentina. ibianco@ceprocor.uncor.edu
authors.forename: Ismael D
authors.initials: ID
authors.lastname: Bianco
authors.forename: José J
authors.initials: JJ
authors.lastname: Daniele
authors.forename: Cristina
authors.initials: C
authors.lastname: Delgado
authors.forename: Derek
authors.initials: D
authors.lastname: Fisher
authors.forename: Gillian E
authors.initials: GE
authors.lastname: Francis
authors.forename: Gerardo D
authors.initials: GD
authors.lastname: Fidelio
mesh_terms: D000818:Animals; D017837:Bothrops; D002850:Chromatography, Gel; D003435:Crotalid Venoms; D004546:Elapid Venoms; D017815:Elapidae; D006031:Glycosylation; D007700:Kinetics; D010741:Phospholipases A / Q000302:isolation & purification / Q000378:metabolism*; D054467:Phospholipases A2; D011092:Polyethylene Glycols; D013379:Substrate Specificity
chemical_list: D003435:Crotalid Venoms; D004546:Elapid Venoms; D011092:Polyethylene Glycols; D010741:Phospholipases A; D054467:Phospholipases A2