Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan, China.
Proteome Sci. 2010 Jun 18;8:35. doi: 10.1186/1477-5956-8-35.
Proteomic analysis of laticifer latex in Hevea brasiliensis has been received more significant attentions. However, the sticky and viscous characteristic of rubber latex as cytoplasm of laticifer cells and the complication of laticifer latex membrane systems has made it challenge to isolate high-quality proteins for 2-DE and MS.
Based on the reported Borax/PVPP/Phenol (BPP) protocol, we developed an efficient method for protein preparation from different latex subcellular fractions and constructed high-resolution reference 2-DE maps. The obtained proteins from both total latex and C-serum fraction with this protocol generate more than one thousand protein spots and several hundreds of protein spots from rubber particles as well as lutoid fraction and its membranes on the CBB stained 2-DE gels. The identification of 13 representative proteins on 2-DE gels by MALDI TOF/TOF MS/MS suggested that this method is compatible with MS.
The proteins extracted by this method are compatible with 2-DE and MS. This protein preparation protocol is expected to be used in future comparative proteomic analysis for natural rubber latex.
巴西橡胶树乳汁的蛋白质组分析受到了更多的关注。然而，胶乳作为乳管细胞细胞质的粘性和粘性特征以及乳管乳胶膜系统的复杂性，使得分离高质量的 2-DE 和 MS 蛋白质具有挑战性。
基于已报道的硼砂/PVPP/酚（BPP）方案，我们开发了一种从不同乳胶亚细胞部分制备蛋白质的有效方法，并构建了高分辨率参考 2-DE 图谱。从总胶乳和 C 血清部分获得的蛋白质与该方案生成了一千多个蛋白质斑点，以及从橡胶颗粒以及lutoid 部分及其膜上获得的几百个蛋白质斑点在 CBB 染色的 2-DE 凝胶上。通过 MALDI TOF/TOF MS/MS 对 2-DE 凝胶上的 13 个代表性蛋白质的鉴定表明，该方法与 MS 兼容。
该方法提取的蛋白质与 2-DE 和 MS 兼容。该蛋白质制备方案有望用于未来天然橡胶乳的比较蛋白质组学分析。
