Purification and characterization of the biological effects of phospholipase A(2) from sea anemone Bunodosoma caissarum.
Sea anemones contain a variety of biologically active substances. Bunodosoma caissarum is a sea anemone from the Cnidaria phylum, found only in Brazilian coastal waters. The aim of the present work was to study the biological effects of PLA(2) isolated from the sea anemone B. caissarum on the isolated perfused kidney, the arteriolar mesenteric bed and on insulin secretion. Specimens of B. caissarum were collected from the São Vicente Channel on the southern coast of the State of São Paulo, Brazil. Reverse phase HPLC analysis of the crude extract of B. caissarum detected three PLA(2) proteins (named BcPLA(2)1, BcPLA(2)2 and BcPLA(2)3) found to be active in B. caissarum extracts. MALDI-TOF mass spectrometry of BcPLA(2)1 showed one main peak at 14.7 kDa. The N-terminal amino acid sequence of BcPLA(2)1 showed high amino acid sequence identity with PLA(2) group III protein isolated from the Mexican lizard (PA23 HELSU, HELSU, PA22 HELSU) and with the honey bee Apis mellifera (PLA(2) and 1POC_A). In addition, BcPLA(2)1 also showed significant overall homology to bee PLA(2). The enzymatic activity induced by native BcPLA(2)1 (20 microg/well) was reduced by chemical treatment with p-bromophenacyl bromide (p-BPB) and with morin. BcPLA(2)1 strongly induced insulin secretion in presence of high glucose concentration. In isolated kidney, the PLA(2) from B. caissarum increased the perfusion pressure, renal vascular resistance, urinary flow, glomerular filtration rate, and sodium, potassium and chloride levels of excretion. BcPLA(2)1, however, did not increase the perfusion pressure on the mesenteric vascular bed. In conclusion, PLA(2), a group III phospholipase isolated from the sea anemone B. caissarum, exerted effects on renal function and induced insulin secretion in conditions of high glucose concentration.
Martins
René D
RD
Department of Physiology and Pharmacology - Institute of Biomedicine and Clinical Research Unit - Federal University of Ceará, Fortaleza, CEP-60.420-970, Ceará, Brazil.
Alves
Renata S
RS
Martins
Alice M C
AM
Barbosa
Paulo Sergio F
PS
Evangelista
Janaina S A M
JS
Evangelista
João José F
JJ
Ximenes
Rafael M
RM
Toyama
Marcos H
MH
Toyama
Daniela O
DO
Souza
Alex Jardelino F
AJ
Orts
Diego J B
DJ
Marangoni
Sérgio
S
de Menezes
Dalgimar B
DB
Fonteles
Manassés C
MC
Monteiro
Helena S A
HS
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D005947:Glucose / Q000378:metabolism; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007668:Kidney / Q000187:drug effects; D008297:Male; D008638:Mesenteric Arteries / Q000187:drug effects; D008969:Molecular Sequence Data; D054467:Phospholipases A2 / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D012615:Sea Anemones / Q000201:enzymology*; D016415:Sequence Alignment; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D007328:Insulin; D054467:Phospholipases A2; D005947:Glucose