聚糖
光解
化学
糖蛋白
紫外线
色谱法
鉴定(生物学)
亲水作用色谱法
紫外线辐射
质谱法
生物化学
高效液相色谱法
光化学
生物
材料科学
植物
光电子学
放射化学
作者
Virginia K. James,Annika A.M. van der Zon,Edwin E. Escobar,Sean D. Dunham,Andrea Gargano,Jennifer S. Brodbelt
标识
DOI:10.1021/acs.jproteome.4c00600
摘要
Protein glycosylation is implicated in a wide array of diseases, yet glycoprotein analysis remains elusive owing to the extreme heterogeneity of glycans, including microheterogeneity of some of the glycosites (amino acid residues). Various mass spectrometry (MS) strategies have proven tremendously successful for localizing and identifying glycans, typically utilizing a bottom-up workflow in which glycoproteins are digested to create glycopeptides to facilitate analysis. An emerging alternative is top-down MS that aims to characterize intact glycoproteins to allow precise identification and localization of glycans. The most comprehensive characterization of intact glycoproteins requires integration of a suitable separation method and high performance tandem mass spectrometry to provide both protein sequence information and glycosite localization. Here, we couple ultraviolet photodissociation and hydrophilic interaction chromatography with high resolution mass spectrometry to advance the characterization of intact glycoproteins ranging from 15 to 34 kDa, offering site localization of glycans, providing sequence coverages up to 93%, and affording relative quantitation of individual glycoforms.
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