双向BN/SDS-PAGE分离鉴定痢疾杆菌福氏5型M90T株毒力相关膜蛋白复合物  

Identification of Virulence-related Membrane Protein Complex from Shigella flexneri 5a M90T by BN/SDS-PAGE

作　　者：龚益熊[1] 廖翔[2] 朱力[2] 吴兰[1] 周晓巍[2] 梁龙[2] 黄培堂[2] 

机构地区：[1]南昌大学食品工程与生命科学学院,南昌330047 [2]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通报》2011年第9期163-166,180,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(30970122)

摘　　要：通过蓝色非变性凝胶电泳(BN-PAGE)比较痢疾杆菌福氏5型野生株M90T和大质粒缺失株M90TΔT的膜蛋白复合物,发现一个野生株特有的复合物,其分子量约为290 kD,命名为M90T-290。通过第二向SDS-PAGE分离M90T-290得到6个蛋白亚基,质谱鉴定为:一个由大质粒编码的毒力蛋白Apyrase(ATP-二磷酸水解酶)和5个染色体编码的蛋白,这些蛋白可能以膜复合物的形式影响毒力蛋白IcsA的单极性分布和痢疾杆菌在细胞间的扩散。这个新发现的毒力相关膜复合物在痢疾杆菌的致病过程中可能发挥重要作用。Virulence-related membrane protein complexes is very important for Shigella＇s pathogenesis,such as type III secretion system.Here we describe an efficient method to isolate membrane protein complexes from S.flexneri 5a wild type strain M90T and its large plasmid cured mutant M90TΔT.A wild type specific band was found in complexes samples of M90T by Blue Native PAGE（BN-PAGE）separation and comparison,which is about 290 kD and so named M90T-290.M90T-290 was then dissociated into subunits by second dimensional SDS-PAGE and identified by MALDI-TOF MS.Six subunit proteins were identified,including a virulence large plasmid encoded protein（apyrase）and several chromosome encoded proteins.These proteins may formed a novel membrane complex which is involved in unipolar distribution of IcsA and intracellular spread of Shigella,and play an important role in pathogenesis of Shigella.

关 键 词：痢疾杆菌 蓝色非变性凝胶电泳 膜蛋白复合物 毒力 ATP-二磷酸水解酶 

分 类 号：R378[医药卫生—病原生物学]