细菌蛋白质组双向电泳中“串联珠”现象初探  

Protein spot train in 2-DE gels for bacterial proteome:a primary research

作　　者：胡威[1,2] 朱力[2] 商娜[2] 王雪芳[2] 徐慧杰[2] 冯尔玲[2] 魏华[1] 王恒樑[1,2] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,南昌330047 [2]军事医学科学院生物工程研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《军事医学》2011年第1期48-53,共6页Military Medical Sciences

基　　金：国家自然科学基金项目(30700035);国家973计划项目(2005CB522904)

摘　　要：目的初步探讨细菌样品双向电泳中"串联珠"状蛋白点产生的原因。方法通过对比不同样品制备方法对双向电泳胶图的影响,分析"串联珠"状蛋白点出现的条件,并取"串联珠"状蛋白点进行胶内酶解及MALDI-TOF/TOF分析,寻找"串联珠"状蛋白点可能产生的原因。结果以含0.5%SDS的裂解液煮沸菌体的方法制备样品,就会产生"串联珠"状蛋白点。蛋白鉴定结果显示"串联珠"状蛋白点为同一蛋白,MALDI-TOF/TOF一级质谱峰没有出现明显的偏移峰。结论 "串联珠"产生的原因是由于SDS在加热条件下会对蛋白产生修饰或紧密结合作用,特别是相对分子质量较大的酸性蛋白。Objective To find the possible reason for making spot train in 2-DE gels of bacterial proteome.Methods The 2-DE gels with different sample preparation methods were compared to discover the key factor for making spot train.The spots forming a train were cut off form the gels and digested by trypsin,then analyzed by MALDI-TOF/TOF.Results Spot train would appear when the protein samples were boiled in lysis buffer containing 0.5% SDS.The spots on one train were identified to be the same protein by MALDI-TOF/TOF.No significant shift of the mass peaks was observed in the PMF spectra of these protein spots.Conclusion SDS would lead to a certain modification or tight binding to proteins when boiled,especially those acidic proteins with large relative molecular mass.

关 键 词：双向电泳 基质辅助激光解吸电离飞行时间质谱 串联珠 十二烷硫酸钠 

分 类 号：Q51[生物学—生物化学] R378[医药卫生—病原生物学]