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本文旨在分析Xcc 8004菌株分泌蛋白的蛋白表达谱,建立Xcc 8004分泌谱分析方法。首先将诱导的Xcc 8004分泌蛋白进行液体酶解,通过HPLC阳离子交换柱分离多肽,采用EASY-nLC结合Orbitrap质谱鉴定分泌谱。经过过滤筛选后,共精确鉴定蛋白1 532个,SignalP分析其中包含286个含信号肽且不含跨膜区,其中118个为假定蛋白。对286个可能的分泌蛋白进行基因本体评注分析,其中,主要为行驶水解酶类,受体蛋白,信号传导等重要的微生物-植物互作介导蛋白。本研究通过液相色谱-质谱联用能够高效鉴定Xcc 8004的分泌蛋白,对这些鉴定的分泌蛋白的信息学分析有利于我们进一步研究Xcc 8004的致病机制,为研究微生物植物相互作用鉴定了方法与理论基础。
The purpose of this article is to analyze the protein expression profile of the secreted protein of strain Xcc 8004, and to establish a method for the analysis of the secretion profile of Xcc 8004. The induced Xcc 8004 secreted protein was first subjected to liquid enzymatic digestion, and the polypeptide was separated by HPLC cation exchange column. The secretion profile was identified by EASY-nLC combined with Orbitrap mass spectrometry. After filtering and screening, a total of 1 532 proteins were accurately identified. SignalP analysis contained 286 signal peptides and contained no transmembrane domain, of which 118 were putative proteins. Genome-wide commentary analyzes of 286 possible secretory proteins were performed, of which the major microbial-plant interaction mediators are hydrolysates, receptor proteins, and signal transduction. In this study, the secreted proteins of Xcc 8004 can be efficiently identified by LC-MS, and the informative analysis of these identified secreted proteins is helpful for us to further study the pathogenesis of Xcc 8004 and to identify methods for studying microbial interactions And theoretical basis.