Exploring roles of the chitinase ChiC in modulating Pseudomonas aeruginosa virulence phenotypes

Proteins were precipitated using chloroform and methanol, by the protocol described by Wessel and Flügge (1984). Following precipitation, the proteins were reduced and alkylated before being digested, cleaned and desalted using STrap tips (Zougman A, Selby PJ, Banks RE. 2014). The peptide samples were analyzed by a nano UPLC (nanoElute, Bruker) coupled to a trapped ion mobility spectrometry and a quadrupole time of flight mass spectrometer (timsTOF Pro, Bruker).

采用Wessel与Flügge于1984年发表的实验方法，使用氯仿与甲醇对蛋白质进行沉淀。沉淀完成后，先对蛋白质进行还原与烷基化处理，随后采用STrap 吸头（STrap tips）完成酶解、净化与脱盐操作（Zougman A、Selby PJ、Banks RE，2014）。肽段样品通过纳流超高效液相色谱仪（nanoElute，布鲁克）连接至捕集离子淌度-四极杆飞行时间联用质谱仪（timsTOF Pro，布鲁克）完成分析检测。