Research and Development Center, Ezequiel Dias Foundation, 30510-010, Belo Horizonte, Brazil.
J Proteomics. 2012 Apr 3;75(7):2181-95. doi: 10.1016/j.jprot.2012.01.020. Epub 2012 Jan 24.
We report the comparative proteomic characterization of the venoms of Bothrops atrox, B. barnetti and B. pictus. The venoms were subjected to RP-HPLC and the resulting fractions analyzed by SDS-PAGE. The proteins were cut from the gels, digested with trypsin and identified via peptide mass fingerprint and manual sequencing of selected peptides by MALDI-TOF/TOF mass spectrometry. Around 20-25 proteins were identified belonging to only 6-7 protein families. Metalloproteinases of the classes P-I and P-III were the most abundant proteins in all venoms (58-74% based on peak area A214 nm), followed by phospholipases-A(2) (6.4-14%), disintegrins (3.2-9%) and serine proteinases (7-11%), and some of these proteins occurred in several isoforms. In contrast cysteine-rich secretory proteins and L-amino acid oxidases appeared only as single isoforms and were found only in B. atrox and B. barnetti. C-type lectins were also detected in all venoms but at low levels (~ 5%). Furthermore, the venoms contain variable numbers of peptides (<3 kDa) and non-protein compounds which were not considered in this work. The protein composition of the investigated Bothrops species is in agreement with their pharmacological and pathological effects.
我们报告了矛头蝮蛇（Bothrops atrox）、巴塔戈尼亚矛头蝮蛇（B. barnetti）和巴西矛头蝮蛇（B. pictus）毒液的比较蛋白质组学特征。毒液经过 RP-HPLC 分离，得到的馏分用 SDS-PAGE 进行分析。将凝胶中的蛋白质切下，用胰蛋白酶消化，通过 MALDI-TOF/TOF 质谱仪的肽质量指纹图谱和选定肽段的手动测序进行鉴定。大约有 20-25 种蛋白质被鉴定，仅属于 6-7 种蛋白质家族。所有毒液中含量最丰富的蛋白质是 P-I 和 P-III 类金属蛋白酶（基于 214nm 吸光度峰面积的 58-74%），其次是磷脂酶 A2（6.4-14%）、分裂素（3.2-9%）和丝氨酸蛋白酶（7-11%），其中一些蛋白质存在多个同工型。相比之下，富含半胱氨酸的分泌蛋白和 L-氨基酸氧化酶仅以单个同工型出现，仅存在于矛头蝮蛇（B. atrox）和巴塔戈尼亚矛头蝮蛇（B. barnetti）中。C 型凝集素也存在于所有毒液中，但含量较低（约 5%）。此外，毒液中还含有数量不定的（<3 kDa）肽和非蛋白质化合物，本研究未对其进行考虑。所研究的矛头蝮蛇种的蛋白质组成与其药理学和病理学效应一致。
