Liao D J, Lu X P, Chen H S, Lu Y, Mo Z Y
State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
Chinese Academy of Tropical Agricultural Sciences Environment and Plant Protection Institute, Danzhou, China.
Genet Mol Res. 2016 Aug 30;15(3):gmr9006. doi: 10.4238/gmr.15039006.
The peel of mango (Mangifera indica L.) is a special plant tissue that contains many compounds that interfere with protein extraction. A successful separation with Two-dimensional electrophoresis (2-DE) is the key step for proteomic analysis. To evaluate the efficiencies of mango peel protein extraction for 2-DE, four extraction methods were tested: 1) 2-D clean-up kit, 2) trichloroacetic acid/acetone precipitation, 3) phenol extraction, 4) phenol with methanol/ammonium acetate precipitation. The results showed that the phenol with methanol/ammonium acetate precipitation produced the best quality protein extraction and separation. Proteins were separated in 30-70 and >70 kDa ranges better than with the other methods. Acidic proteins had better resolution with fewer horizontal and vertical streaks. Sixteen proteins were identified by maxtrix-assisted laser desorption/ ionisation time-of-flight tandem mass spectrometry (MALDI-TOF/ TOF-MS/MS). The result demonstrated that each of these four methods can be used to prepare mango peel proteins. The phenol with methanol/ ammonium acetate precipitation was the best choice for proteomic analysis of mango peel.
芒果（Mangifera indica L.）果皮是一种特殊的植物组织，含有许多干扰蛋白质提取的化合物。二维电泳（2-DE）成功分离是蛋白质组学分析的关键步骤。为了评估用于二维电泳的芒果果皮蛋白质提取效率，测试了四种提取方法：1）二维清理试剂盒；2）三氯乙酸/丙酮沉淀法；3）酚提取法；4）酚与甲醇/醋酸铵沉淀法。结果表明，酚与甲醇/醋酸铵沉淀法产生的蛋白质提取和分离质量最佳。在30 - 70 kDa和>70 kDa范围内，蛋白质的分离效果比其他方法更好。酸性蛋白质具有更好的分辨率，水平和垂直条纹更少。通过基质辅助激光解吸/电离飞行时间串联质谱（MALDI-TOF/TOF-MS/MS）鉴定了16种蛋白质。结果表明，这四种方法中的每一种都可用于制备芒果果皮蛋白质。酚与甲醇/醋酸铵沉淀法是芒果果皮蛋白质组学分析的最佳选择。
