Evidence that a 16-kilodalton integral membrane protein antigen from Schistosoma japonicum adult worms is a type A2 phospholipase

Abstract

Type A2 phospholipase (PLA2) activity has been observed in integral membrane protein extracts of Schistosoma japonicum. Antiserum raised against bee venom PLA2 recognized a single 16-kDa band in the parasite extracts; it also localized to antigen in the gut lining of fixed adult schistosomes as shown by immunofluorescence techniques. Evidence was obtained that the molecule was expressed at low levels in comparison with other integral membrane proteins and was weakly immunogenic in rabbits. Two oligonucleotide probes were constructed on the basis of highly conserved regions between the nucleotide sequences of rat, bovine, rattlesnake, and bee venom PLA2; these probes were used to isolate S. japonicum genomic DNA phage clones. A 1.8-kb FnuD2 fragment was shown by Southern blot analysis to strongly hybridize with the 5' 32P-labeled PLA2 oligonucleotides in both S. japonicum genomic DNA and DNA from one of the phage clones. The nucleotide and predicted amino acid sequences of this fragment revealed homology with the C terminus of PLA2s from different species.