Simultaneous analysis of cellular glycoproteome and phosphoproteome in cervical carcinoma by one-pot specific...

Cervical carcinoma is a kind of common gynecological cancer, the physiopathology of which is consanguineously connected with protein glycosylation and phosphorylation. Here, the magnetic binary metal oxide composites with the functionalization of hydrophilic tripeptide is designed and synthesized for simultaneous enrichment of glycopeptide and phosphopeptide in HeLa cells. Multiple kinds of metal sites realize more comprehensive enrichment of phosphopeptides compared with single metal, and ultra-hydrophilic property of glutathione further endows the composites with excellent affinity towards glycopeptides. The composites exhibited low detection limit (0.5 fmol/μL) and high selectivity (digests of β-casein and BSA, 1:500, m/m) for phosphopeptides, also showed good enrichment efficiency for glycopeptide including detection limit (0.1 fmol/μL) and selectivity (digests of HRP and BSA, 1:50, m/m). Meanwhile, the composites still possess outstanding enrichment capability towards phosphopeptide and glycopeptide after stored for two months or six consecutive times reuse. Eventually, 1177 phosphopeptides and 438 glycopeptides are identified simultaneously from 100 μg HeLa cell digests. As anticipated, potential biomarkers, such as heat shock protein beta-1, DNA topoisomerase 2-alpha, proliferation marker protein Ki-67 and 60 kDa heat shock protein are detected, suggesting its promising application in discovery and screening of cervical carcinoma.