Rapid Serum Proteome Profiling of Glycation and N-Glycosylation in Women with a History of Gestational Diabetes

Here, we developed and validated a high throughput proteomics workflow that enables combined profiling of global proteomes and post-translational modifications (PTMs) without conventional enrichment or fractionation. Using mass spectrometry-based methods, we identified PTM landscapes, particularly N-glycopeptides and non-enzymatic glycated peptides, from the same sample prepared for global proteomics. Applying this methodology to investigate longitudinal serum in women at 6 and 10 years post-gestational diabetes mellitus (GDM), we identified PTM signatures, including N-glycosylation and non-enzymatic glycation, associated with development of type 2 diabetes (T2D) and prediabetes, which were confirmed in an independent T2D cohort.