Citation: WANG Shu-Yue, JIA Sheng-Nan, LI Xiao-Ying, ZHANG Hai-Yang, FENG Xin, WANG Shi, TAO Wei-Guo, HU Liang-Hai. A Preliminary Study of Human Urine Exosome Glycoproteome[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(10): 1334-1342. doi: 10.19756/j.issn.0253-3820.201220 shu

A Preliminary Study of Human Urine Exosome Glycoproteome

1.
School of Life Sciences, Jilin University, Changchun 130023, China;
2.
The Second Hospital of Jilin University, Changchun 130041, China;
3.
School of Biological Science & Medical Engineering, Southeast University, Nanjing 210096, China;
4.
Purdue University, West Lafayette, IN 47907

Received Date: 21 April 2020
Revised Date: 6 July 2020

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 21675061).

Protein glycosylation of human urine exosome can reflect important biological information, which promotes a large-scale identification for the study of urine exosome glycoproteome. In this work, a two step enrichment method based on extracellular vesicles total recovery and purification (EVTRAP) and click maltose materials was developed for urine glycoproteome study. Based on the lipid bilayer of exosome, EVTRAP, a lipophilic magnetic beads, could effectively extract exosomes from human urine. After lysis and digestion of exosomes, based on the hydrophilic glycan chains of glycopeptides, glycopeptides were effectively enriched by click maltose hydrophilic materials. The exosomes proteins digestion and glycopeptides were analyzed by LC-MS-MS. Finally, 1925 nonredundant proteins and 14233 peptides were obtained at proteome level, which indicated that the urine exosomes were extracted with highly specificity. Meanwhile, 468 nonredundant intact glycopeptides from 88 glycoproteins corresponding to 95 various glycans on 135 glycosites were identified. Furthermore, the glycopeptides with high heterogeneity and different glycan structures were analyzed. This study provided an effective approach for investigation of urine exosome glycoproteome for disease biomarker discovery.
- Exosome,
- Glycosylation,
- Urine,
- Extracellular vesicles total recovery and purification,
- Click maltose
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沈阳化工大学材料科学与工程学院沈阳 110142