Fractionation and proteomic analysis of the Walterinnesia aegyptia snake venom using OFFGEL and MALDI-TOF-MS techniques
Fractionation and proteomic analysis of the Walterinnesia aegyptia snake venom using OFFGEL and MALDI-TOF-MS techniques
复制标题
DOI：
10.1002/elps.201500207
复制
发表时间：
2015-10-01
期刊：
影响因子：
2.9
通讯作者：
De Waard, Michel
中科院分区：
文献类型：
--
作者：
El Aziz, Tarek Mohamed Abd;Bourgoin-Voillard, Sandrine;De Waard, Michel
动物毒液是100多种不同化合物的复杂混合物，包括多肽、蛋白质和非蛋白质化合物，如脂类、碳水化合物和金属离子。此外，现有的化合物在这些毒液中显示出广泛的相对分子质量和浓度，使得分离和纯化程序相当繁琐。在这里，我们首次用MS分析了OFFGEL技术用于埃及斑潜蝇毒液成分分离的优势，并与SEC和RP-HPLC两种经典的分离方法进行了比较。我们证明，OFFGEL在更大范围的馏分中分离毒液成分，保持关于给定化合物在相邻馏分中存在的适当分辨率，并允许MS识别更多数量的离子(总计134个离子，102个离子)。我们还得出结论，在离子检测方面，应用几种分离技术(SEC和RP-HPLC以及OFFGEL)可以提供互补的结果(SEC多21个，RP-HPLC多22个)。因此，我们结合使用所有这些技术，提供了埃及斑潜蝇毒液中存在的134种离子的完整清单。
Animal venoms are complex mixtures of more than 100 different compounds, including peptides, proteins, and nonprotein compounds such as lipids, carbohydrates, and metal ions. In addition, the existing compounds show a wide range of molecular weights and concentrations within these venoms, making separation and purification procedures quite tedious. Here, we analyzed for the first time by MS the advantages of using the OFFGEL technique in the separation of the venom components of the Egyptian Elapidae Walterinnesia aegyptia snake compared to two classical methods of separation, SEC and RP-HPLC. We demonstrate that OFFGEL separates venom components over a larger scale of fractions, preserve respectable resolution with regard to the presence of a given compound in adjacent fractions and allows the identification of a greater number of ions by MS (102 over 134 total ions). We also conclude that applying several separating techniques (SEC and RP-HPLC in addition to OFFGEL) provides complementary results in terms of ion detection (21 more for SEC and 22 more with RP-HPLC). As a result, we provide a complete list of 134 ions present in the venom of W. aegyptia by using all these techniques combined.