cDNA cloning and sequencing of phospholipase A2 from the pyloric ceca of the starfish Asterina pectinifera.
Three cDNA from the pyloric ceca of the starfish Asterina pectinifera, (namely, cDNA 1, 2, and 3), encoding phospholipase A2 (PLA2), were isolated and sequenced. These cDNAs were composed of 415 bp with an open reading frame of 414 bp at nucleotide positions 1-414, which encodes 138 amino acids including N-terminal Met derived from the PCR primer. The amino acid sequence deduced from the cDNA 1 was completely consistent with the sequence determined with the starfish PLA2 protein, while those deduced from cDNA 2 and cDNA 3 differed at one and twelve amino acid residual positions, respectively, from the sequence of the PLA2 protein, suggesting the presence of multiple forms in the starfish PLA2. All of the sequences deduced from cDNA 1, 2, and 3 required two amino acid deletions in pancreatic loop region, and sixteen insertions and three deletions in beta-wing region when aligned with the sequence of mammalian pancreatic PLA2. In phylogenetic tree, the starfish PLA2 should be classified into an independent group, but hardly to the established groups IA and IB. The characteristic structure in the pancreatic loop and beta-wing regions may account for the specific properties of the starfish PLA2, e.g. the higher activity and characteristic substrate specificity compared with commercially available PLA2 from porcine pancreas.
Kishimura
H
H
Department of Marine Bioresources Chemistry, Faculty of Fisheries, Hokkaido University, Hakodate, Japan. kishi@fish.hokudai.ac.jp
Ojima
T
T
Hayashi
K
K
Nishita
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002432:Cecum / Q000201:enzymology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000737:chemistry / Q000235:genetics*; D054467:Phospholipases A2; D010802:Phylogeny; D016415:Sequence Alignment; D013215:Starfish / Q000201:enzymology / Q000235:genetics*
D018076:DNA, Complementary; D010741:Phospholipases A; D054467:Phospholipases A2