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首页> 外文期刊>Journal of Molecular Biology >Sequencing and two-dimensional structure prediction of a phospholipase A(2) inhibitor from the serum of the common tiger snake (Notechis scutatus)
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Sequencing and two-dimensional structure prediction of a phospholipase A(2) inhibitor from the serum of the common tiger snake (Notechis scutatus)

机译：从普通老虎蛇（Notechis scutatus）的血清中的磷脂酶A（2）抑制剂的测序和二维结构预测

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摘要

A phospholipase A, 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. 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.
机译：在普通虎蛇（Notechis scutatus）的血清中已鉴定出磷脂酶A抑制剂。该抑制剂由两条链组成，一条α链和一条β链形成非共价结合的复合物，该复合物能够抑制被测试的所有磷脂酶A.酶的酶活性。 α和β链已纯化至均一，已消化并测序。根据产生的肽序列，设计简并的PCR引物，并使用5'和3'RACE PCR阐明链的完整cDNA序列。总共鉴定出三种α-链同工型，仅检测到一种β-链同工型。使用五个预测程序预测了3条α链和1条β链的二维结构（区分二级结构类别;最邻近的二级结构;海德堡的轮廓网络;基于多重比对的自我优化的预测方法; SSPAL） 。对于每个蛋白质链，产生共识预测。讨论了有关蛋白质功能的结果，以及它们如何影响抑制剂的三维结构。此外，几种蛇磷脂酶A（2）抑制剂的序列被用作基序预测算法（MEME）的输入。讨论了有关这些蛋白质活性的结果。

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