Venoms of Russell's vipers (genus Daboia) are known for their deadly coagulopathic and other effects. We herein studied various isoforms of venom phospholipases A(2) (PLAs) from two Daboia species at their geographic boundary. From Myanmar Daboia siamensis venom (designated as DsM), four PLAs (designated DsM-aI, aI', aII' and bI') were purified, and the cDNAs encoding two acidic (DsM-aI and aII) and two basic PLAs (DsM-bI and S1) were also cloned from its venom-glands. DsM-S1 is identical to the major venom PLA of southern India Daboia russelii, but the protein is absent from the venom. Additionally, four PLAs (designated DrK-aI, aII, bI and bII) were cloned from cDNA obtained from venom glands of a Kolkata D. russelii, and the PLAs were purified from the pooled venom (designated as DrK). The acidic DrK-aI is the most neurotoxic and lethal among these PLAs; DsM-aI which differs from DrK-aI by only the Phe2 substitution shows greatly reduced enzymatic activity and lethality. Both acidic PLAs do not form dimeric complex with basic PLAs in the same venoms. DsM-bI' is neurotoxic and lethal but its orthologous DrK-bI (97% identical to DsM-bI') is a much weaker toxin. Given the fact that most of the orthologous PLAs of DrK and DsM share 97-100% sequence identity, Daboia vipers of Myanmar and Kolkata must be closely related. Molecular phylogenetic analyses on 30 venom PLAs of Eurasian vipers' revealed co-evolution of five subtypes of venom PLAs in both Daboia and Vipera genera. Our results shed light on the intra- and inter-species variations and structure-function relationships of viperid venom PLAs.

译文

:Russell的毒蛇(Daboia属)的毒液以致命的凝血病和其他功效而闻名。我们在这里研究了来自两个Daboia物种在其地理边界处的毒性磷脂酶A(2)(PLA)的各种同工型。从缅甸Daboia siamensis毒液(称为DsM)中,纯化了四个PLA(分别称为DsM-aI,aI',aII'和bI'),并编码了两个酸性(DsM-aI和aII)和两个碱性PLA(DsM)的cDNA。 -bI和S1)也从其毒腺中克隆得到。 DsM-S1与印度南部Daboia russelii的主要毒液PLA相同,但该毒液中不存在该蛋白质。另外,从得自罗氏加尔各答蛇毒腺的cDNA克隆了四个PLA(命名为DrK-aI,aII,bI和bII),并从合并的毒液(命名为DrK)中纯化了PLA。在这些PLA中,酸性DrK-aI具有最高的神经毒性和致死性。 DsM-aI与DrK-aI的不同之处仅在于Phe2取代,显示酶活性和杀伤力大大降低。两种酸性PLA在同一毒液中均不会与碱性PLA形成二聚体复合物。 DsM-bI'具有神经毒性和致死性,但其直系同源的DrK-bI(与DsM-bI'97%相同)是一种弱得多的毒素。鉴于大多数DrK和DsM的直系同源PLA拥有97-100%的序列同一性,因此缅甸和加尔各答的Daboia蛇蝎必须紧密相关。对欧亚vi蛇的30种毒液PLA进行分子系统发育分析,揭示了Daboia和Vipera属中5种亚型毒液PLA的共同进化。我们的研究结果揭示了蛇毒PLA的种内和种间变异以及结构与功能的关系。

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