Quantitative Longitudinal Inventory of the N-Glycoproteome of Human Milk from a Single Donor Reveals the Highly Variable Repertoire and Dynamic Site-Specific Changes.

摘要: Protein N-glycosylation on human milk proteins assists in protecting an infant's health and functions among others as competitive inhibitors of pathogen binding immunomodulators. Due to the individual uniqueness each mother's overall complexity temporal changes protein N-glycosylation, analysis N-glycoproteome requires longitudinal personalized approaches, providing protein- N-site-specific quantitative information. Here, we describe automated platform using hydrophilic-interaction chromatography (HILIC)-based cartridges enabling proteome-wide monitoring intact N-glycopeptides just a digest 150 μg breast protein. We were able map around 1700 glycopeptides from 110 glycoproteins covering 191 glycosites, which 43 sites have not been previously reported with experimental evidence. next quantified 287 these originating 50 targeted proteomics approach. Although glycoprotein, site, attached glycan revealed distinct dynamic changes, did observe few general trends. For instance, fucosylation, especially terminal increased across lactation period. Building improved glycoproteomics approach outlined above, future studies are warranted reveal potential impact observed glycosylation microheterogeneity healthy development infants.