Enzymatic and structural characterization of a basic phospholipase A(2) from the sea anemone Condylactis gigantea.
This work aimed at the isolation and structural/functional characterization of a phospholipase A(2) (CgPLA(2)) from the extract of the anemone Condylactis gigantea. CgPLA(2) was isolated with a high purity level through three chromatographic steps, showing pI 8.6 and molecular weights of 14,500 and 29,000 for the monomer and dimer, respectively. CgPLA(2) showed a high catalytic activity upon fluorescent phospholipids inducing no direct hemolytic activity. This enzyme, which is Ca(2+)-dependent, showed a lower stability against temperature and pH variations when compared with snake venom enzymes. The enzymatic activity was significantly reduced or completely abolished after chemical modification of CgPLA(2) with BPB. Its cDNA was then obtained, with 357 base pairs which codified for a mature protein of 119 amino acid residues. A comparative analysis of the primary structure of CgPLA(2) revealed 84%, 61%, 43% and 42% similarity to the PLA(2)s from Adamsia carciniopados, Nematostella vectensis, Vipera russelli russelli and Bothrops jararacussu, respectively.
Romero
Lazara
L
Centro de Estudios de Proteínas, Facultad de Biología, Universidad de La Habana, Cuba.
Marcussi
Silvana
S
Marchi-Salvador
Daniela P
DP
Silva
Floriano P
FP
Fuly
André L
AL
Stábeli
Rodrigo G
RG
da Silva
Saulo L
SL
González
Jorge
J
Monte
Alberto Del
AD
Soares
Andreimar M
AM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002853:Chromatography, Liquid; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D006461:Hemolysis; D006863:Hydrogen-Ion Concentration; D008297:Male; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D054467:Phospholipases A2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D012615:Sea Anemones / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D054467:Phospholipases A2