{"doi":"10.1016/j.mcpro.2021.100081","title":"In-depth Site-specific Analysis of N-glycoproteome in Human Cerebrospinal Fluid and Glycosylation Landscape Changes in Alzheimer's Disease","abstract":"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.","journal":"Molecular & Cellular Proteomics","year":2021,"id":151748,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":80,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.8674,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":588186,"name":"Qinying Yu","orcid":"0000-0001-9483-1478","position":1,"is_corresponding":false},{"id":228749,"name":"Qing Yu","orcid":"0000-0003-0468-5353","position":2,"is_corresponding":false},{"id":645606,"name":"Jillian Johnson","orcid":null,"position":3,"is_corresponding":false},{"id":644823,"name":"Richard David Shipman","orcid":"0000-0003-4731-5563","position":4,"is_corresponding":false},{"id":2381,"name":"Xiaofang Zhong","orcid":"0000-0001-8831-5733","position":5,"is_corresponding":false},{"id":348903,"name":"Junfeng Huang","orcid":"0000-0002-1672-3751","position":6,"is_corresponding":false},{"id":289695,"name":"Sanjay Asthana","orcid":"0000-0002-3998-4479","position":7,"is_corresponding":false},{"id":289694,"name":"Cynthia M. Carlsson","orcid":"0000-0001-5743-1563","position":8,"is_corresponding":false},{"id":493820,"name":"Ozioma C. Okonkwo","orcid":"0000-0001-5803-6055","position":9,"is_corresponding":false},{"id":289711,"name":"Lingjun Li","orcid":"0000-0003-0056-3869","position":10,"is_corresponding":false},{"id":348905,"name":"Zhengwei Chen","orcid":"0000-0002-5900-3359","position":0,"is_corresponding":true}],"reference_count":160,"raw_metadata":null,"created_at":"2026-07-18T23:43:16.823357Z","pmid":"33862227","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}