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
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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