志贺菌敏感株与诱导耐多药株蛋白质组学分析

Analysis on different proteomics between sensitive strain and induced anti-multidrug strain of Shigella flexneri

目的对志贺菌敏感株与诱导耐多药株全菌蛋白进行蛋白质组学比较,寻找细菌耐多药相关蛋白。方法采用4类抗生素的次抑菌浓度,对临床分离鉴定的志贺菌敏感株进行诱导耐多药试验;对志贺菌敏感株及诱导耐多药株全菌蛋白进行双向电泳;电泳图谱采用Image Master 2D Platinum软件分析;差异表达蛋白进行基质辅助激光解吸飞行时间质谱(MOLDI TOF-TOF质谱)分析。结果成功获得志贺菌诱导耐多药株,在志贺菌敏感株与耐多药株全菌蛋白质图谱中分别检测出(946±37)个和(1 096±189)个蛋白质斑点;共发现48个差异表达的蛋白点,其中5个质谱鉴定为ABC转运蛋白、谷胺酰转肽酶、天冬氨酸氨甲酰基转移酶、翻译延长因子Tu、单链DNA结合蛋白;其中前3个蛋白中在耐多药株中表达量增强,后2个减弱。结论5个耐多药相关蛋白中在细胞代谢中起重要作用的酶类表达量上调;ABC转运蛋白在志贺菌诱导耐多药机制中起重要作用。

Objective To search for the new proteins related to multidrug resistance, the proteomics of whole cellular proteins between sensitive strain and induced anti - multidrug strain of shigella flexneri by using of two- dimensional electrophoresis（2 - DE） were compared. Methods Clinical sensitive shigella flexneri strain was induced into anti - drug strain by 1/2 MIC induced trials of four kinds of antibiotic; Immobilized pH gradient（IPG） two- dimensional electrophoresis was adopted and the gels were analyze by Image Master 2D Platinum software; Matrix assisted laser desorption/ionization- time of flight mass spectrometry（MALDI - TOF MS） were used to analyze differential expression proteins. Results Induced anti -multidrug strain of shigella flexneri was obtained successfully. It was found that there were （946 ± 37） protein spots in whole cellular protein 2 - DE gels of sensitive strain and （1 096 ±189） protein spots in that of induced anti - multidrug strain, 48 differential expression protein spots were found and 5 proteins related to multidrug resistance were identified based on peptide mass fingerprinting： ATP binding cassette transporter protein, γ- glutamyltranspeptidase, aspartate carbamoyl- ransferase were high expression in induced anti- multidrug strain;But chain elongation factor EF- Tu and SSB were high expression in sensitive strain. Conclusion By mass spectrum analysis of 5 proteins related to multidrug resistance, some important cellular metabolism enzymes expressed highly, ATP binding cassette transporter protein affected seriously the mechanism of shigella flexneri multidrug resistance.