Global proteomic and functional analysis of Crotalus durissus collilineatus individual venom variation and its impact on envenoming
Global proteomic and functional analysis of Crotalus durissus collilineatus individual venom variation and its impact on envenoming
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DOI：
10.1016/j.jprot.2018.02.020
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发表时间：
2019-01-16
影响因子：
3.3
通讯作者：
Arantes, Eliane Candiani
中科院分区：
文献类型：
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作者：
de Oliveira, Isadora Sousa;Cardoso, Iara Aime;Arantes, Eliane Candiani
个体差异研究对于了解蛇咬伤中毒并提高抗蛇毒血清的产生及其有效性非常重要。因此，本研究的目的是通过蛋白质组学技术对 22 个 Crotalus durissus collilineatus 标本的毒液成分的种内变异进行比较分析。通过 RP-FPLC 分离毒液，并通过 SDS-PAGE 和质谱分析。尽管相似，但色谱和电泳图谱显示出显着的定性和定量差异。首次在C. d collilineatus中鉴定出一些毒液成分，如谷胱甘肽过氧化物酶、神经生长因子、5'-核苷酸酶、血管紧张素转换酶、羧肽酶、磷酸二酯酶、谷氨酰胺环化酶和磷脂酶B。关于透明质酸酶活性，2种毒液在测试量中未检测到酶活性。此外，小鼠体内的响尾蛇毒液表明，不同标本的毒液会导致生化和免疫学参数（例如肌酸激酶和白细胞介素6）的多样化变化。这项研究表明，响尾蛇的毒液存在显着的种内差异。 collilineatus，这可能会影响抗蛇毒血清治疗的产生和有效性。生物学意义：本研究对 22 C. d. 进行了蛋白质组和功能分析。对 collilineatus 个体的毒液进行了分析，并验证了它们之间发生的质和量的变化。所评估的毒液引起的毒液具有不同的生化和免疫学参数变化。这些结果证实，在制备抗蛇毒血清时需要使用具有最大可能变异性的毒液库，以提高其有效性。此外，这项研究首次鉴定出该亚种毒液中的 8 种不同蛋白质，增加了对其成分的了解，并表明它是这些蛋白质的来源，并具有可能的生物技术应用。
Individual variations studies are important to understand the snakebite envenoming and to improve the anti-venom production and its effectiveness. In this way, the objective of this study was a comparative analysis of intraspecific variation in the venom composition of 22 Crotalus durissus collilineatus specimens through proteomic techniques. Venoms were fractionated by RP-FPLC, and analyzed by SDS-PAGE and mass spectrometry. Although similar, chromatographic and electrophoretic profiles showed significant qualitative and quantitative differences. Some venom components were identified for the very first time in C. d collilineatus, such as glutathione peroxidase, nerve growth factor, 5'-nucleotidase, angiotensin-converting enzyme, carboxypeptidase, phosphodiesterase, glutaminyl cyclase and phospholipase B. Regarding hyaluronidase activity, 2 venoms did not present detectable enzyme activity in the tested amounts. Additionally, in vivo crotalic envenoming in mice showed that venoms from different specimens resulted in diversified changes of biochemical and immunological parameters, such as creatine kinase and interleukin 6. This study demonstrated significant intraspecific variations in the venom of C. d. collilineatus, which may impact the production and effectiveness of the antivenom therapy.Biological significance: This study performed the proteomic and functional analyzes of 22 C. d. collilineatus individual venoms and verified the occurrence of quali and quantitative variations among them. The venoms evaluated caused envenomings with different changes in biochemical and immunological parameters. These results confirm the need to use a pool of venoms with the greatest possible variability in the preparation of antivenoms, in order to improve their effectiveness. In addition, this study was able to identify for the first time 8 different proteins in this subspecies venom, increasing knowledge about its composition and showing that it is a source of these proteins with possible biotechnological applications.