cDNA cloning of the two subunits of phospholipase A2 inhibitor PLIgamma from blood plasma of the Chinese mamushi, Agkistrodon blomhoffii siniticus.
Three phospholipase A2 (PLA2) inhibitors (PLI) have been purified from the blood plasma of the Chinese mamushi, Agkistrodon blomhoffii siniticus; 1 of these, PLIgamma, contains 2 homologous subunits, PLIgamma-A and PLIgamma-B. The cDNAs encoding these 2 subunits of PLIgamma were isolated from a liver cDNA library by using fragments from polymerase chain reaction amplifications as probes and sequenced. The respective nucleotide sequences encoded 19-residue signal sequences, followed by 181-residue proteins. The calculated molecular masses were 20123 and 20150 Da for the PLIgamma-A and PLIgamma-B subunits, respectively; and PLIgamma-A included a N-linked carbohydrate site at Asn-157. The sequences of these subunits contained 2 internal repeats of disulfide-bonding pattern characteristic to those of urokinase-type plasminogen activator receptor and members of the Ly-6 superfamily. A phylogenetic analysis comparing the amino acid sequences of PLIgamma-A and PLIgamma-B with those for other snakes revealed that the gene duplication leading to these 2 subunits occurred before the divergence of Viperidae and Elapidae.
Okumura
K
K
Department of Biochemistry, Osaka University of Pharmaceutical Sciences, Takatsuki, Japan.
Inoue
S
S
Ohkura
N
N
Ikeda
K
K
Hayashi
K
K
D017836:Agkistrodon / Q000097:blood* / Q000145:classification / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary; D004220:Disulfides / Q000737:chemistry; D015723:Gene Library; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010726:Phosphodiesterase Inhibitors / Q000097:blood*; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D054467:Phospholipases A2; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D020449:Repetitive Sequences, Amino Acid; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D004220:Disulfides; D006023:Glycoproteins; D046911:Macromolecular Substances; D010726:Phosphodiesterase Inhibitors; C090541:phospholipase A2 inhibitory protein, Naja naja kaouthia; D010741:Phospholipases A; D054467:Phospholipases A2