弗氏志贺菌磷酸化蛋白的检测

Detection of Phosphorylated Proteins of Shigella flexneri

目的:检测弗氏志贺菌全菌蛋白中被磷酸化修饰的蛋白。方法:制取弗氏2a志贺菌2457T野生株全菌磷酸化蛋白样品时加入磷酸化酶抑制剂,随后对样品进行双向电泳,以抗磷酸丝氨酸/苏氨酸/酪氨酸抗体为免疫探针,通过Western印迹找到被磷酸化的蛋白,并进行胶内酶解及MALDI-TOF质谱分析。结果与结论:共检测到13个磷酸化蛋白,其中9个为代谢途径中的酶。

Objective： To detect phosphorylated proteins of Shigella flexneri.Methods： Whole-cell proteins of wild-type S.flexneri 2a 2457T treated with phosphatase inhibitor were separated by two-dimensional electrophoresis.One of the gels was used for Western blotting to detect phosphorylated proteins.The corresponding immunogenicity spots found on the other Coomassie brilliant blue stained gel were cut out,digested by trypsin and analyzed by MALDI-TOF MS.Results Conclusion： Thirteen phosphorylated proteins were detected,and 9 of them are enzymes involved in metabolic pathway.