earticle

영어

Protein post-translational modification (PTM) increases the functional diversity of the proteome by the covalent addition of proteins. Therefore, identifying and understanding PTMs is critical in the study of protein bioscience. Protein glycosylation is acknowledged as one of the major post -translational modifications, with significant effects on protein folding, conformation, stability and activity. The need to characterize glycoproteins continues to increase as more irrefutable examples of the essential role that covalent carbohydrates m the proteins play in biological processes and functions. The extant challenge of glycobiology and glycotechnology is to approach the extremely low level of sensitivity, used for PTM analysis over the protein identification. In this study, several biologically important proteomic samples was analyzed by shotgun proteomics method, the resulting modified peptides are separated and detected by ultrahigh -performance nano liquid chromatography-tandem mass spectrometry (nano LC-MS/MS). We, for the first time, identified specific 0-GleN Ac modification sites of Octl, Oct4, and 0-linked glycosylation site of Tango 1 with the state-of-the-art nano LC-Orbitrap Fusion Tribrid mass spectrometer. Additionally, We developed the Metabolic Isotope Labeling of Polysaccharides with Isotopic Glucose (MILPIG) method to label the light C2C) or heavy CuC1) glucose on glycans of plant, yeast, and fungi. we quantitatively analyzed the N-linked glycan of Oryza sativa (rice) usmg metabolic labeling with stable isotopic glucose by a mass spectrometry. We also applied metabolic isotope labeling strategy for glycan quantitation in yeast (Saccharomyces cerevisiae).