Venomic and Transcriptomic Analysis of Centipede Scolopendra subspinipes dehaani
Venomic and Transcriptomic Analysis of Centipede Scolopendra subspinipes dehaani
复制标题
蜈蚣蜈蚣的毒液和转录组学分析
DOI：
10.1021/pr300881d
复制
发表时间：
2012-12-01
影响因子：
4.4
通讯作者：
Zhang, Yun
中科院分区：
文献类型：
--
作者：
Liu, Zi-Chao;Zhang, Rong;Zhang, Yun
蜈蚣的第一对肢上有毒腺，其毒液中含有大量具有不同生化和药理特性的成分。然而，有关其毒液的成分和功能的信息在很大程度上是未知的。在这项研究中，通过转录组学和蛋白质组学分析以及生物功能测定，系统地研究了蜈蚣毒液。在随机筛选大约 1500 个独立克隆后，使用来自单个蜈蚣物种的一对毒腺从构建的 cDNA 文库中克隆了 1122 个全长 cDNA 序列，编码 543 种不同的蛋白质。转录组分析显示，神经毒素、离子通道作用成分和毒液过敏原是粗毒液的主要成分。同时，从S. subspinipes dehaani的粗毒液中纯化并表征了40种蛋白质/肽。这40种蛋白质或肽中有29种的N端氨基酸测序和质谱结果与其相应的cDNA匹配良好。纯化的蛋白/肽表现出不同的药理特性，包括：（1）血小板聚集活性； (2)抗凝血活性； (3)磷脂酶A(2)活性； (4)胰蛋白酶抑制活性； (5)电压门控钾通道活性； (6)电压门控钠通道活性； (7)电压门控钙通道活动。其中大多数与已知公共数据库中存储的其他蛋白质序列没有显着相似性。这项工作提供了数量最多的具有医学药学意义的候选蛋白质或肽，并揭示了蜈蚣S. subspinipes dehaani毒液的毒素性质。
Centipedes have venom glands in their first pair of limbs, and their venoms contain a large number of components with different biochemical and pharmacological properties. However, information about the compositions and functions of their venoms is largely unknown. In this study, Scolopendra subspinipes dehaani venoms were systematically investigated by transcriptomic and proteomic analysis coupled with biological function assays. After random screening approximately 1500 independent clones, 1122 full length cDNA sequences, which encode 543 different proteins, were cloned from a constructed cDNA library using a pair of venom glands from a single centipede species. Neurotoxins, ion channel acting components and venom allergens were the main fractions of the crude venom as revealed by transcriptomic analysis. Meanwhile, 40 proteins/peptides were purified and characterized from crude venom of S. subspinipes dehaani. The N-terminal amino acid sequencing and mass spectrum results of 29 out of these 40 proteins or peptides matched well With their corresponding cDNAs. The purified proteins/peptides showed different pharmacological properties, including the following: (1) platelet aggregating activity; (2) anticoagulant activity; (3) phospholipase A(2) activity; (4) trypsin inhibiting activity; (5) voltage gated potassium channel activities; (6) voltage gated sodium channel activities; (7) voltage gated calcium channel activities. Most of them showed no significant similarity to other protein sequences deposited in the known public database. This work provides the largest number of protein or peptide candidates with medical-pharmaceutical significance and reveals the toxin nature of centipede S. subspinipes dehaani venom.