Sequencing and two-dimensional structure prediction of a phospholipase A(2) inhibitor from the serum of the common tiger snake (Notechis scutatus).
A phospholipase A(2) inhibitor has been identified in the serum of the common tiger snake (Notechis scutatus). The inhibitor is composed of two chains, an alpha-chain and a beta-chain, that form a non-covalently associated complex capable of inhibiting the enzymatic activity of all phospholipase A(2) enzymes it was tested against. The alpha and beta-chains have been purified 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 5' and 3' RACE PCR. A total of three alpha-chain isoforms were identified, only one isoform of the beta-chain was detected. The two-dimensional structure of the three alpha-chains and one beta-chain were predicted using five prediction programs (discrimination of secondary structure class; nearest neighbour secondary structure, profile 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 influence the three-dimensional structure of the inhibitor. Additionally, the sequences of several snake phospholipase A(2) inhibitors were used as the input for a motif prediction algorithm (MEME). The results are discussed in relation to the activity of these proteins.
Hains
P G
PG
Department of Cell and Molecular Biology, Toxin Research Group, University of Technology, Sydney Westbourne St, Gore Hill 2065, Australia.
Nield
B
B
Sekuloski
S
S
Dunn
R
R
Broady
K
K
D000478:Alkylation; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001798:Blood Proteins / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D004220:Disulfides / Q000378:metabolism; D017815:Elapidae / Q000097:blood / Q000235:genetics; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D017421:Sequence Analysis
D001798:Blood Proteins; D004220:Disulfides; C400481:PLIalpha; D010741:Phospholipases A