Recombinant O-GlcNAcylated human tau after GalNAzylation, biotinylation, trypsinization, coupling to streptavidin beads and β-elimination-Michael addition - RAW data files - nLC-Q Exactive HF acquisition

Chemo-enzymatic derivatization (GalNAzylation, biotinylation) of O-GlcNAcylated recombinant human tau, trypsinization, enrichment via coupling to streptavidin beads, and release of tagged peptides by β-elimination – Michael addition reaction. nLC-HRMS (Q Exactive HF) analysis of final extract to monitor sulfited peptides for the localization of tau O-GlcNAcylation sites. Data were acquired in data dependent MS2 (ddMS2) mode and for low abundant O-GlcNAc peptides also in parallel reaction monitoring (PRM) mode.

通过化学酶促衍生化（包括GalNAzylation和生物素化）处理O-GlcNAc基化的重组人tau蛋白，进行胰蛋白酶解，通过与亲和素磁珠耦合进行富集，并通过β-消除-迈克尔加成反应释放标记肽。使用nLC-HRMS（Q Exactive HF）分析最终提取物，以监测硫代肽的定位，从而确定tau蛋白的O-GlcNAcylation位点。数据采集采用数据依赖的MS2（ddMS2）模式，对于低丰度的O-GlcNAc肽，同时采用并行反应监测（PRM）模式。