Decoding Protein Glycosylation by an Integrative Mass Spectrometry-Based De Novo Sequencing Strategy Article Swipe
Jing Gao
,
Hong‐Xu Chen
,
Hongrui Yin
,
Xin Chen
,
Zhicheng Yang
,
Yuqiu Wang
,
Jianhong Wu
,
Yinping Tian
,
Hong Shao
,
Liuqing Wen
,
Hu Zhou
·
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1021/jacsau.4c00960
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1021/jacsau.4c00960
Glycoproteins, representing more than 50% of human proteins and most biopharmaceuticals, are crucial for regulating various biological processes. The complexity of multiple glycosylation sites often leads to incomplete sequence coverage and ambiguous glycan modification profiles. Here, we developed an integrative mass spectrometry-based approach for decoding unknown glycoproteins, which is featured with the combination of deglycosylation-mediated de novo sequencing with glycosylation site characterization. We utilized the enzymatic deglycosylation of N-/ O-glycans to achieve comprehensive sequence coverage. Additionally, EThcD fragmentation enables the identification of high-quality long peptides, facilitating precise protein assembly. We subsequently applied this method to de novo sequencing of the highly glycosylated therapeutic fusion protein Etanercept (Enbrel). We also sequenced three new tumor necrosis factor receptor:Fc-fusion biologics with largely unknown sequences, unveiling subtle distinctions in the primary sequences. Furthermore, we characterized N- and O-glycosylation modifications of these proteins at subunit, glycopeptide, and glycan levels. This strategy bridges the gap between the de novo sequencing and glycosylation modification, providing comprehensive information on the primary structure and glycosylation modifications for glycoproteins. Notably, our method could be a robust solution for accurate sequencing of the glycoproteins and has practical value not only in basic research but also in the biopharmaceutical industry.
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- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1021/jacsau.4c00960
- OA Status
- gold
- Cited By
- 1
- References
- 43
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4406715125
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- OpenAlex ID
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https://openalex.org/W4406715125Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1021/jacsau.4c00960Digital Object Identifier
- Title
-
Decoding Protein Glycosylation by an Integrative Mass Spectrometry-Based De Novo Sequencing StrategyWork title
- Type
-
articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2025Year of publication
- Publication date
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2025-01-22Full publication date if available
- Authors
-
Jing Gao, Hong‐Xu Chen, Hongrui Yin, Xin Chen, Zhicheng Yang, Yuqiu Wang, Jianhong Wu, Yinping Tian, Hong Shao, Liuqing Wen, Hu ZhouList of authors in order
- Landing page
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https://doi.org/10.1021/jacsau.4c00960Publisher landing page
- Open access
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YesWhether a free full text is available
- OA status
-
goldOpen access status per OpenAlex
- OA URL
-
https://doi.org/10.1021/jacsau.4c00960Direct OA link when available
- Concepts
-
Glycosylation, Mass spectrometry, Computational biology, Chemistry, Decoding methods, Biology, Biochemistry, Computer science, Chromatography, AlgorithmTop concepts (fields/topics) attached by OpenAlex
- Cited by
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1Total citation count in OpenAlex
- Citations by year (recent)
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2025: 1Per-year citation counts (last 5 years)
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43Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.coverage. | 74 |
| abstract_inverted_index.developed | 37 |
| abstract_inverted_index.enzymatic | 65 |
| abstract_inverted_index.industry. | 197 |
| abstract_inverted_index.peptides, | 84 |
| abstract_inverted_index.practical | 185 |
| abstract_inverted_index.profiles. | 34 |
| abstract_inverted_index.providing | 157 |
| abstract_inverted_index.sequenced | 109 |
| abstract_inverted_index.structure | 163 |
| abstract_inverted_index.unveiling | 121 |
| abstract_inverted_index.<i>N-</i>/ | 68 |
| abstract_inverted_index.Etanercept | 105 |
| abstract_inverted_index.biological | 16 |
| abstract_inverted_index.complexity | 19 |
| abstract_inverted_index.incomplete | 27 |
| abstract_inverted_index.processes. | 17 |
| abstract_inverted_index.regulating | 14 |
| abstract_inverted_index.sequences, | 120 |
| abstract_inverted_index.sequences. | 127 |
| abstract_inverted_index.sequencing | 57, 97, 153, 179 |
| abstract_inverted_index.combination | 52 |
| abstract_inverted_index.information | 159 |
| abstract_inverted_index.integrative | 39 |
| abstract_inverted_index.therapeutic | 102 |
| abstract_inverted_index.Furthermore, | 128 |
| abstract_inverted_index.distinctions | 123 |
| abstract_inverted_index.facilitating | 85 |
| abstract_inverted_index.glycosylated | 101 |
| abstract_inverted_index.high-quality | 82 |
| abstract_inverted_index.modification | 33 |
| abstract_inverted_index.representing | 1 |
| abstract_inverted_index.subsequently | 90 |
| abstract_inverted_index.Additionally, | 75 |
| abstract_inverted_index.characterized | 130 |
| abstract_inverted_index.comprehensive | 72, 158 |
| abstract_inverted_index.fragmentation | 77 |
| abstract_inverted_index.glycopeptide, | 140 |
| abstract_inverted_index.glycoproteins | 182 |
| abstract_inverted_index.glycosylation | 22, 59, 155, 165 |
| abstract_inverted_index.modification, | 156 |
| abstract_inverted_index.modifications | 134, 166 |
| abstract_inverted_index.Glycoproteins, | 0 |
| abstract_inverted_index.glycoproteins, | 46 |
| abstract_inverted_index.glycoproteins. | 168 |
| abstract_inverted_index.identification | 80 |
| abstract_inverted_index.deglycosylation | 66 |
| abstract_inverted_index.<i>O-</i>glycans | 69 |
| abstract_inverted_index.biopharmaceutical | 196 |
| abstract_inverted_index.characterization. | 61 |
| abstract_inverted_index.receptor:Fc-fusion | 115 |
| abstract_inverted_index.spectrometry-based | 41 |
| abstract_inverted_index.biopharmaceuticals, | 10 |
| abstract_inverted_index.<i>O-</i>glycosylation | 133 |
| abstract_inverted_index.deglycosylation-mediated | 54 |
| cited_by_percentile_year.max | 95 |
| cited_by_percentile_year.min | 91 |
| corresponding_author_ids | https://openalex.org/A5085026154, https://openalex.org/A5045675715 |
| countries_distinct_count | 1 |
| institutions_distinct_count | 11 |
| corresponding_institution_ids | https://openalex.org/I131434179, https://openalex.org/I19820366, https://openalex.org/I4210112820, https://openalex.org/I4210165038 |
| citation_normalized_percentile.value | 0.75204574 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | True |