期刊
ACS CHEMICAL BIOLOGY
卷 17, 期 11, 页码 3059-3068出版社
AMER CHEMICAL SOC
DOI: 10.1021/acschembio.1c00932
关键词
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资金
- National Institutes of Health (NIH) [RF1 AG052324, R21AG065728, U01CA231081, R01 DK071801, P30AG062715]
- NIH [NIH-NCRR S10RR029531]
- Office of the Vice Chancellor for Research and Graduate Education at the University of Wisconsin-Madison
- Wisconsin Alumni Research Foundation
- University of Wisconsin-Madison School of Pharmacy
Site-specific O-glycoproteome mapping in complex biological systems provides a molecular basis for understanding the structure-function relationships of glycoproteins and their roles in physiological and pathological processes. A comprehensive strategy was developed to analyze the O-glycoproteome in cerebrospinal fluid (CSF) and identify O-glycopeptides from both sialylated and nonsialylated glycoforms. The study also revealed that endogenous peptides in CSF were predominantly O-glycosylated, with a decreasing trend in fucosylation and an increasing trend in endogenous peptide O-glycosylation observed in patients with Alzheimer's disease.
Site-specific O-glycoproteome mapping in complex biological systems provides a molecular basis for understanding the structure-function relationships of glycoproteins and their roles in physiological and pathological processes. Previous O-glycoproteome analysis in cerebrospinal fluid (CSF) focused on sialylated glycoforms, while missing information on other glycosylation types. In order to achieve an unbiased O-glycosylation profile, we developed an integrated strategy combining universal boronic acid enrichment, high-pH fractionation, and electron-transfer and higher-energy collision dissociation (EThcD) for enhanced intact O-glycopeptide analysis. We applied this strategy to analyze the Oglycoproteome in CSF, resulting in the identification of 308 O-glycopeptides from 110 O-glycoproteins, covering both sialylated and nonsialylated glycoforms. To our knowledge, this is the largest data set of O-glycoproteins and O-glycosites reported for CSF to date. We also developed a peptidomics workflow that utilized the EThcD and a three-step database searching strategy for comprehensive PTM analysis of endogenous peptides, including N-glycosylation, O-glycosylation, and other common peptide PTMs. Interestingly, among the 1411 endogenous peptides identified, 89 were O-glycosylated, and only one N-glycosylated peptide was found, indicating that CSF endogenous peptides were predominantly O-glycosylated. Analyses of the O-glycoproteome and endogenous peptidome PTMs were also conducted in the CSF of MCI and AD patients to provide a landscape of glycosylation patterns in different disease states. Our results showed a decreasing trend in fucosylation and an increasing trend of endogenous peptide O-glycosylation, which may play an important role in AD progression.
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