Sequencing and Two-Dimensional Structure Prediction of a Phospholipase A2 Inhibitor from the Serum of the Common Tiger Snake

Academic Press Ltd
Publication Type:
Journal Article
Journal of Molecular Biology, 2001, 312 pp. 875 - 884
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A phospholipase A2 inhibitor has been identi®ed in the serum of the common tiger snake (Notechis scutatus). The inhibitor is composed of two chains, an a-chain and a b-chain, that form a non-covalently associated complex capable of inhibiting the enzymatic activity of all phospholipase A2 enzymes it was tested against. The a and b-chains have been puri®ed to homogeneity, digested and sequenced. From the peptide sequence generated, degenerate PCR primers were designed and used to elucidate the complete cDNA sequence of the chains using 50 and 30 RACE PCR. A total of three a-chain isoforms were identi®ed, only one isoform of the b-chain was detected. The two-dimensional structure of the three a-chains and one b-chain were predicted using ®ve prediction programs (discrimination of secondary structure class; nearest neighbour secondary structure, pro®le network from Heidelberg; self-optimised prediction method from multiple alignment, SSPAL). For each protein chain a consensus prediction was generated. Results are discussed in relation to the function of the protein, and how they may in¯uence the three-dimensional structure of the inhibitor. Additionally, the sequences of several snake phospholipase A2 inhibitors were used as the input for a motif prediction algorithm (MEME). The results are discussed in relation to the activity of these proteins.
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