Preparation of O-Glycopeptides from commercial bovine whey proteins using offline liquid chromatography-Mass spectrometry.

Preparation of O-Glycopeptides from commercial bovine whey proteins using offline liquid chromatography-Mass spectrometry. Carbohydr Res. 2020 Mar 19;491:107981 Authors: Kurogochi M, Matsuda A, Mizuno M Abstract O-Glycopeptides derived from natural bioresources are an attractive material for a variety of purposes. Whey protein products are used as a human dietary supplement and in animal feed and are a readily available resource for the preparation of O-glycopeptides. The protein composition of bovine milk is well-studied, and many glycoproteins carrying N-glycans and O-glycans have been found in commercial whey protein products. In particular, κ-casein glycomacropeptide and lactophorin, which have several O-glycans, are known to exist in whey protein. Here, we report an isolation method of O-glycopeptides bearing disialyl core 1 type and core 2 type glycan moieties from commercially available whey protein products using proteose peptone extraction, enzymatic digestion (with trypsin or thermolysin), and sequential high-performance liquid chromatography purification. We were able to isolate several kinds of O-glycopeptides from lactophorin and κ-casein: six peptide sequences and five kinds of O-glycans. The O-glycopeptides were detected and identified by flow injection analysis combined with electrospray ionization mass spectrometry and tandem mass spectrometry using collision-induced dissociation and electron transfer dissociation....
Source: Carbohydrate Research - Category: Genetics & Stem Cells Authors: Tags: Carbohydr Res Source Type: research