In-depth Site-specific Analysis of N-glycoproteome in Human Cerebrospinal Fluid and Glycosylation Landscape Changes in Alzheimer's Disease
| Autor: | Qing Yu, Jillian Johnson, Qinying Yu, Junfeng Huang, Cynthia M. Carlsson, Xiaofang Zhong, Sanjay Asthana, Lingjun Li, Richard David Shipman, Ozioma C. Okonkwo, Zhengwei Chen |
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| Rok vydání: | 2020 |
| Předmět: |
Special Issue: Glycoproteomics
Glycosylation Proteome ConA concanavalin A Disease DMSO dimethyl sulfoxide Biochemistry CSF cerebrospinal fluid Analytical Chemistry Pathogenesis electron-transfer higher-energy collision induced dissociation (EThcD) chemistry.chemical_compound Cerebrospinal fluid ADRC Alzheimer’s Disease Research Center Extracellular fluid Amyloid precursor protein Fucosylation ABC ammonium bicarbonate chemistry.chemical_classification 0303 health sciences biology 030302 biochemistry & molecular biology Glycopeptides glycopeptide enrichment RCA Ricinus communis agglutinin AD Alzheimer's disease SDS sodium dodecyl sulfate FDR false discovery rate WGA wheat germ agglutinin Computational biology CNS central nervous system cerebrospinal fluid Cell Line TFA trifluoroacetic acid 03 medical and health sciences HILIC hydrophilic interaction chromatography Alzheimer Disease APP amyloid precursor protein Humans Molecular Biology N-glycoproteome analysis 030304 developmental biology Glycoproteins PBA phenylboronic acid Research chemistry DTT dithiothreitol site-specific intact glycopeptide characterization biology.protein EThcD electron transfer and higher-energy collision dissociation PTM posttranslational modification Glycoprotein IAA iodoacetamide |
| Zdroj: | Molecular & Cellular Proteomics : MCP |
| ISSN: | 1535-9484 |
| Popis: | As the body fluid that directly interchanges with the extracellular fluid of the central nervous system (CNS), cerebrospinal fluid (CSF) serves as a rich source for CNS-related disease biomarker discovery. Extensive proteome profiling has been conducted for CSF, but studies aimed at unraveling site-specific CSF N-glycoproteome are lacking. Initial efforts into site-specific N-glycoproteomics study in CSF yield limited coverage, hindering further experimental design of glycosylation-based disease biomarker discovery in CSF. In the present study, we have developed an N-glycoproteomic approach that combines enhanced N-glycopeptide sequential enrichment by hydrophilic interaction chromatography (HILIC) and boronic acid enrichment with electron transfer and higher-energy collision dissociation (EThcD) for large-scale intact N-glycopeptide analysis. The application of the developed approach to the analyses of human CSF samples enabled identifications of a total of 2893 intact N-glycopeptides from 511 N-glycosites and 285 N-glycoproteins. To our knowledge, this is the largest site-specific N-glycoproteome dataset reported for CSF to date. Such dataset provides molecular basis for a better understanding of the structure–function relationships of glycoproteins and their roles in CNS-related physiological and pathological processes. As accumulating evidence suggests that defects in glycosylation are involved in Alzheimer's disease (AD) pathogenesis, in the present study, a comparative in-depth N-glycoproteomic analysis was conducted for CSF samples from healthy control and AD patients, which yielded a comparable N-glycoproteome coverage but a distinct expression pattern for different categories of glycoforms, such as decreased fucosylation in AD CSF samples. Altered glycosylation patterns were detected for a number of N-glycoproteins including alpha-1-antichymotrypsin, ephrin-A3 and carnosinase CN1 etc., which serve as potentially interesting targets for further glycosylation-based AD study and may eventually lead to molecular elucidation of the role of glycosylation in AD progression. Graphical abstract Highlights • Efficient N-glycopeptide sequential enrichment by HILIC and boronic acid enrichment. • Site-specific intact N-glycopeptide characterization using EThcD. • In-depth site-specific N-glycoproteome analysis in human CSF. • Mapping the landscape of glycosylation patterns in Alzheimer's disease. In Brief An exploratory glycosylation-based biomarker study has been conducted for in-depth mapping of an overall glycosylation landscape and site-specific alteration in glycoproteome collected from cerebrospinal fluids (CSF) in healthy control and Alzheimer’s disease (AD) subjects. The comparison will shed light on the glycoproteome profile, dominant glycosylation differences and similarities, and some of the interesting glycoprotein candidates with specific glycosylation pattern alterations in AD. |
| Databáze: | OpenAIRE |
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