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Analysis of evolution and phylogeny of snakes of Trimeresurus genus in the southwestern islands of Japan based on structural information of venom-gland isozymes

基于毒腺同工酶结构信息的日本西南诸岛竹叶青属蛇的进化与系统发育分析

基本信息

批准号：
15570089
负责人：
OHNO Motonori
金额：
$ 2.05万
依托单位：
Sojo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

1. Interisland mutation of Trimeresurus flavoviridis venom phospholipase A_2 (PLA_2) isozymes.The venoms of T.flavoviridis snakes from Amami-Oshima, Tokunoshima and Okinawa islands contain basic [Asp^<49>]PLA_2s called PLA-B', PLA-B and PL-Y, respectively. They have 1-4 amino acid substitutions that are considered due to occurrence of interisland mutation under isolated environments. A neurotoxic PLA_2 isolated from Amami-Oshima T.flavoviridis venom was named PLA-N. Tokunoshima T.flavoviridis venom also contained this isozyme but Okinawa T.flavoviridis venom contains one amino acid-substituted homolog, called PLA-N(O). This is also a example of interisland mutation. It is speculated that Okiawa island first splitted from the original island and then the rest was disunited into Amami-Oshima and Tokunoshima.2. Evolution and phylogeny of snakes of Trimeresurus genus in the southwestern islands of JapanTwo novel PLA_2 isozymes with Arg at position 49 ([Arg^<49>]PLA_2) were isolated from T.elegans venom and named TeBP(R)-I and TeBP(R)-II. They are polymorphic at position 3, Val for TeBP(R)-I and lle for TeBP(R)-II. A cDNA encoding [Asp49]PLA_2 called TePLA_2 was cloned from its venom gland. A neutral [Asp49]PLA_2 was purified to homogeneity from T.tokarensis venom. However, production of its venom gland cDNA library failed. A phylogenetic tree was constructed for venom PLA_2 isozymes of Trimeresurus genus, except for T.tokarensis, in the southwestern islands of Japan. In the branch of [Lys49]PLA_2-type group, TeBP(R)-I and TeBP(R)-II of T.elegans are much closer to TmK49PLA_2 of T.mucrosquamatus than TfBPI and TfBPI of T.flavoviridis. In PLA2-type group, TePLA_2 is closer to TmPLA_2 than TfPLA2. Thus, T.elegans is the species closer to T.mucrosquamatus than T.flavoviridis. In addition, it is clearly noted that T.okinavensis and T.gramineus are the species fairly distant from T.flavoviridis, T.elegans and T.mucrosquamatus.

1. 竹叶青蛇毒磷脂酶A_2（PLA_2）同工酶的岛间突变。来自奄美大岛、德之岛和冲绳岛的竹叶青蛇毒液中含有碱性[Asp^<49>]PLA_2，分别称为PLA-B'、PLA-B和PL-Y。它们有 1-4 个氨基酸取代，这被认为是由于在隔离环境下发生岛间突变所致。从奄美大岛黄病毒毒液中分离出具有神经毒性的PLA_2，命名为PLA-N。德之岛 T.flavoviridis 毒液也含有这种同工酶，但冲绳 T.flavoviridis 毒液含有一种氨基酸取代的同源物，称为 PLA-N(O)。这也是岛间变异的一个例子。据推测，冲绳岛首先从原来的岛屿中分裂出来，然后其余部分分裂为奄美大岛和德之岛。2．日本西南诸岛竹叶青属蛇的进化与系统发育从竹叶青毒液中分离得到2个49位为Arg的PLA_2同工酶([Arg^<49>]PLA_2)，命名为TeBP(R)-I和TeBP(R)-II。它们在位置 3 处具有多态性，TeBP(R)-I 为 Val，TeBP(R)-II 为 lle。从其毒腺中克隆了编码 [Asp49]PLA_2 的 cDNA（称为 TePLA_2）。从T.tokarensis毒液中纯化出同质的中性[Asp49]PLA_2。然而，其毒腺cDNA文库的生产失败了。对日本西南部诸岛竹叶青属（除托卡伦竹叶青）毒液 PLA_2 同工酶构建了系统发育树。在[Lys49]PLA_2型群分支中，线虫的TeBP(R)-I和TeBP(R)-II比T.flavoviridis的TfBPI和TfBPI更接近T.mucrosquamatus的TmK49PLA_2。在PLA2型组中，TePLA_2比TfPLA2更接近TmPLA_2。因此，T.elegans 是比 T.flavoviridis 更接近 T.mucrosquamatus 的物种。此外，还可以清楚地注意到，T.okinavensis 和 T.gramineus 是与 T.flavoviridis、T.elegans 和 T.mucrosquamatus 相距较远的物种。