title: Purification and renal effects of phospholipase A(2) isolated from Bothrops insularis venom.
abstract: Bothrops insularis venom contains a variety of substances presumably responsible for several pharmacological effects. We investigated the biochemical and biological effects of phospholipase A(2) protein isolated from B. insularis venom and the chromatographic profile showed 7 main fractions and the main phospholipase A(2) (PLA(2)) enzymatic activity was detected in fractions IV and V. Fraction IV was submitted to a new chromatographic procedure on ion exchange chromatography, which allowed the elution of 5 main fractions designated as IV-1 to IV-5, from which IV-4 constituted the main fraction. The molecular homogeneity of this fraction was characterized by high-performance liquid chromatography (HPLC) and demonstrated by mass spectrometry (MS), which showed a molecular mass of 13984.20 Da; its N-terminal sequence presented a high amino acid identity (up to 95%) with the PLA(2) of Bothrops jararaca and Bothrops asper. Phospholipase A(2) isolated from B. insularis (Bi PLA(2) ) venom (10 microg/mL) was also studied as to its effect on the renal function of isolated perfused kidneys of Wistar rats (n=6). Bi PLA(2) increased perfusion pressure (PP), renal vascular resistance (RVR), urinary flow (UF) and glomerular filtration rate (GFR). Sodium (%TNa(+)) and chloride tubular reabsorption (%TCl(-)) decreased at 120 min, without alteration in potassium transport. In conclusion, PLA(2) isolated from B. insularis venom promoted renal alterations in the isolated perfused rat kidney.
authors.affiliation: Department of Pathology, Federal University of Ceara, Fortaleza, Ceará, Brazil.
authors.forename: Marcus Davis
authors.initials: MD
authors.lastname: Machado Braga
authors.forename: Alice Maria
authors.initials: AM
authors.lastname: Costa Martins
authors.forename: Claudênio Diógenes
authors.initials: CD
authors.lastname: Alves
authors.forename: Dalgimar Beserra
authors.initials: DB
authors.lastname: de Menezes
authors.forename: René Duarte
authors.initials: RD
authors.lastname: Martins
authors.forename: Paulo Sérgio
authors.initials: PS
authors.lastname: Ferreira Barbosa
authors.forename: Isadora Maria
authors.initials: IM
authors.lastname: de Sousa Oliveira
authors.forename: Marcos Hikari
authors.initials: MH
authors.lastname: Toyama
authors.forename: Daniela Oliveira
authors.initials: DO
authors.lastname: Toyama
authors.forename: Eduardo Brito
authors.initials: EB
authors.lastname: Dos Santos Diz Filho
authors.forename: Fabio Henrique
authors.initials: FH
authors.lastname: Ramos Fagundes
authors.forename: Manassés Claudino
authors.initials: MC
authors.lastname: Fonteles
authors.forename: Helena Serra
authors.initials: HS
authors.lastname: Azul Monteiro
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops; D003435:Crotalid Venoms / Q000737:chemistry / Q000633:toxicity*; D007684:Kidney Tubules / Q000098:blood supply / Q000187:drug effects*; D008297:Male; D008969:Molecular Sequence Data; D054467:Phospholipases A2 / Q000737:chemistry / Q000633:toxicity*; D051381:Rats; D017208:Rats, Wistar; D012079:Renal Circulation / Q000187:drug effects; D017386:Sequence Homology, Amino Acid
chemical_list: D003435:Crotalid Venoms; C078097:bothropstoxin; D054467:Phospholipases A2