Xu Ning, Zhao Hong-Yan, Yin Yin, Shen Shan-Shan, Shan Lin-Lin, Chen Chuan-Xi, Zhang Yan-Xia, Gao Jian-Fang, Ji Xiang
Hangzhou Key Laboratory for Animal Adaptation and Evolution, College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310036, Zhejiang, China.
Hangzhou Key Laboratory for Animal Adaptation and Evolution, College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310036, Zhejiang, China.
J Proteomics. 2017 Apr 21;159:19-31. doi: 10.1016/j.jprot.2017.02.018. Epub 2017 Mar 3.
We conducted an omics-analysis of the venom of Naja kaouthia from China. Proteomics analysis revealed six protein families [three-finger toxins (3-FTx), phospholipase A (PLA), nerve growth factor, snake venom metalloproteinase (SVMP), cysteine-rich secretory protein and ohanin], and venom-gland transcriptomics analysis revealed 28 protein families from 79 unigenes. 3-FTx (56.5% in proteome/82.0% in transcriptome) and PLA (26.9%/13.6%) were identified as the most abundant families in venom proteome and venom-gland transcriptome. Furthermore, N. kaouthia venom expressed strong lethality (i.p. LD: 0.79μg/g) and myotoxicity (CK: 5939U/l) in mice, and showed notable activity in PLA but weak activity in SVMP, l-amino acid oxidase or 5' nucleotidase. Antivenomic assessment revealed that several venom components (nearly 17.5% of total venom) from N. kaouthia could not be thoroughly immunocaptured by commercial Naja atra antivenom. ELISA analysis revealed that there was no difference in the cross-reaction between N. kaouthia and N. atra venoms against the N. atra antivenom. The use of commercial N. atra antivenom in treatment of snakebites caused by N. kaouthia is reasonable, but design of novel antivenom with the attention on enhancing the immune response of non-immunocaptured components should be encouraged.
The venomics, antivenomics and venom-gland transcriptome of the monocoled cobra (Naja kaouthia) from China have been elucidated. Quantitative and qualitative differences are evident when venom proteomic and venom-gland transcriptomic profiles are compared. Two protein families (3-FTx and PLA) are found to be the predominated components in N. kaouthia venom, and considered as the major players in functional role of venom. Other protein families with relatively low abundance appear to be minor in the functional significance. Antivenomics and ELISA evaluation reveal that the N. kaouthia venom can be effectively immunorecognized by commercial N. atra antivenom, but still a small number of venom components could not be thoroughly immunocaptured. The findings indicate that exploring the precise composition of snake venom should be executed by an integrated omics-approach, and elucidating the venom composition is helpful in understanding composition-function relationships and will facilitate the clinical application of antivenoms.
我们对中国舟山眼镜蛇的毒液进行了组学分析。蛋白质组学分析揭示了六个蛋白质家族[三指毒素（3-FTx）、磷脂酶A（PLA）、神经生长因子、蛇毒金属蛋白酶（SVMP）、富含半胱氨酸的分泌蛋白和ohanin]，毒液腺转录组学分析从79个单基因中揭示了28个蛋白质家族。3-FTx（蛋白质组中占56.5%/转录组中占82.0%）和PLA（26.9%/13.6%）被确定为毒液蛋白质组和毒液腺转录组中最丰富的家族。此外，舟山眼镜蛇毒液在小鼠中表现出很强的致死性（腹腔注射半数致死量：0.79μg/g）和肌毒性（肌酸激酶：5939U/l），并且在PLA方面表现出显著活性，但在SVMP、L-氨基酸氧化酶或5'-核苷酸酶方面活性较弱。抗蛇毒组学评估显示，舟山眼镜蛇的几种毒液成分（占总毒液的近17.5%）不能被市售的中华眼镜蛇抗蛇毒血清完全免疫捕获。酶联免疫吸附测定分析显示，舟山眼镜蛇毒液和中华眼镜蛇毒液与中华眼镜蛇抗蛇毒血清之间的交叉反应没有差异。使用市售的中华眼镜蛇抗蛇毒血清治疗舟山眼镜蛇咬伤是合理的，但应鼓励设计新型抗蛇毒血清，关注增强对未被免疫捕获成分的免疫反应。
已阐明中国舟山眼镜蛇的毒液组学、抗蛇毒组学和毒液腺转录组。比较毒液蛋白质组学和毒液腺转录组学图谱时，定量和定性差异明显。发现两个蛋白质家族（3-FTx和PLA）是舟山眼镜蛇毒液中的主要成分，并被认为是毒液功能作用的主要参与者。其他丰度相对较低的蛋白质家族在功能意义上似乎较小。抗蛇毒组学和酶联免疫吸附测定评估显示，舟山眼镜蛇毒液可被市售的中华眼镜蛇抗蛇毒血清有效免疫识别，但仍有少量毒液成分不能被完全免疫捕获。研究结果表明，应通过综合组学方法探索蛇毒的精确组成，阐明毒液组成有助于理解组成-功能关系，并将促进抗蛇毒血清的临床应用。
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