Purification and renal effects of phospholipase A(2) isolated from Bothrops insularis venom.
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.
Machado Braga
Marcus Davis
MD
Department of Pathology, Federal University of Ceara, Fortaleza, Ceará, Brazil.
Costa Martins
Alice Maria
AM
Alves
Claudênio Diógenes
CD
de Menezes
Dalgimar Beserra
DB
Martins
René Duarte
RD
Ferreira Barbosa
Paulo Sérgio
PS
de Sousa Oliveira
Isadora Maria
IM
Toyama
Marcos Hikari
MH
Toyama
Daniela Oliveira
DO
Dos Santos Diz Filho
Eduardo Brito
EB
Ramos Fagundes
Fabio Henrique
FH
Fonteles
Manassés Claudino
MC
Azul Monteiro
Helena Serra
HS
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
D003435:Crotalid Venoms; C078097:bothropstoxin; D054467:Phospholipases A2