特异性鉴定人细胞中携带 Tn 抗原的糖蛋白。
Specific Identification of Glycoproteins Bearing the Tn Antigen in Human Cells.
机构信息
School of Chemistry and Biochemistry and the Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA, 30332, USA.
出版信息
Angew Chem Int Ed Engl. 2017 Jun 12;56(25):7107-7111. doi: 10.1002/anie.201702191. Epub 2017 May 17.
Abstract
Glycoproteins contain a wealth of valuable information regarding the development and disease status of cells. In cancer cells, some glycans (such as the Tn antigen) are highly up-regulated, but this remains largely unknown for glycoproteins with a particular glycan. Herein, an innovative method combining enzymatic and chemical reactions was first designed to enrich glycoproteins with the Tn antigen. Using synthetic glycopeptides with O-GalNAc (the Tn antigen) or O-GlcNAc, we demonstrated that the method is selective for glycopeptides with O-GalNAc and can distinguish between these two modifications. The diagnostic ions from the tagged O-GalNAc further confirmed the effectiveness of the method and confidence in the identification of glycopeptides with the Tn antigen by mass spectrometry. Using this method, we identified 96 glycoproteins with the Tn antigen in Jurkat cells. The method can be extensively applied in biological and biomedical research.
摘要
糖蛋白包含了大量关于细胞发育和疾病状态的有价值的信息。在癌细胞中，一些聚糖（如 Tn 抗原）被高度上调，但对于具有特定聚糖的糖蛋白，这在很大程度上仍然未知。在此，我们首次设计了一种结合酶和化学反应的创新方法来富集 Tn 抗原的糖蛋白。使用带有 O-GalNAc（Tn 抗原）或 O-GlcNAc 的合成糖肽，我们证明该方法对带有 O-GalNAc 的糖肽具有选择性，并且可以区分这两种修饰。标记的 O-GalNAc 的诊断离子进一步证实了该方法的有效性，并对通过质谱法鉴定 Tn 抗原糖肽的方法有了信心。使用这种方法，我们在 Jurkat 细胞中鉴定出了 96 种带有 Tn 抗原的糖蛋白。该方法可以广泛应用于生物和生物医学研究。
相似文献
1
Specific Identification of Glycoproteins Bearing the Tn Antigen in Human Cells.特异性鉴定人细胞中携带 Tn 抗原的糖蛋白。
Angew Chem Int Ed Engl. 2017 Jun 12;56(25):7107-7111. doi: 10.1002/anie.201702191. Epub 2017 May 17.
2
Simultaneously Identifying and Distinguishing Glycoproteins with O-GlcNAc and O-GalNAc (the Tn Antigen) in Human Cancer Cells.同时鉴定和区分人类癌细胞中的 O-GlcNAc 和 O-GalNAc（Tn 抗原）糖蛋白。
Anal Chem. 2022 Feb 22;94(7):3343-3351. doi: 10.1021/acs.analchem.1c05438. Epub 2022 Feb 8.
3
Mass Spectrometry-Based Chemical and Enzymatic Methods for Global Analysis of Protein Glycosylation.基于质谱的蛋白质糖基化全局分析的化学和酶学方法。
Acc Chem Res. 2018 Aug 21;51(8):1796-1806. doi: 10.1021/acs.accounts.8b00200. Epub 2018 Jul 16.
4
Mass Spectrometric Mapping of Glycoproteins Modified by Tn-Antigen Using Solid-Phase Capture and Enzymatic Release.利用固相捕获和酶释放技术对 Tn 抗原修饰的糖蛋白进行质谱作图。
Anal Chem. 2020 Jul 7;92(13):9230-9238. doi: 10.1021/acs.analchem.0c01564. Epub 2020 Jun 16.
5
Mucin-type O-glycosylation in helminth parasites from major taxonomic groups: evidence for widespread distribution of the Tn antigen (GalNAc-Ser/Thr) and identification of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase activity.主要分类群的蠕虫寄生虫中的粘蛋白型O-糖基化：Tn抗原（GalNAc-Ser/Thr）广泛分布的证据以及UDP-GalNAc：多肽N-乙酰半乳糖胺基转移酶活性的鉴定。
J Parasitol. 2003 Aug;89(4):709-14. doi: 10.1645/GE-2970.
6
COSMC knockdown mediated aberrant O-glycosylation promotes oncogenic properties in pancreatic cancer.COSMC基因敲低介导的异常O-糖基化促进胰腺癌的致癌特性。
Mol Cancer. 2015 May 29;14:109. doi: 10.1186/s12943-015-0386-1.
7
Integrating HexNAcQuest with Glycoproteomics Data Analysis Software to Distinguish HexNAc Isomers on Peptides.将 HexNAcQuest 与糖蛋白质组学数据分析软件集成，以区分肽上的 HexNAc 异构体。
Methods Mol Biol. 2024;2836:67-76. doi: 10.1007/978-1-0716-4007-4_5.
8
Classification of Mucin-Type -Glycopeptides Using Higher-Energy Collisional Dissociation in Mass Spectrometry.使用质谱中的高能量碰撞解离对粘蛋白型糖肽进行分类。
Anal Chem. 2020 Jul 21;92(14):9772-9781. doi: 10.1021/acs.analchem.0c01218. Epub 2020 Jul 9.
9
Unexpected tolerance of glycosylation by UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferase revealed by electron capture dissociation mass spectrometry: carbohydrate as potential protective groups.电子捕获解离质谱揭示 UDP-GalNAc:多肽 α-N-乙酰半乳糖胺基转移酶对糖基化的意外容忍性：碳水化合物作为潜在的保护基团。
Biochemistry. 2010 Jul 20;49(28):5929-41. doi: 10.1021/bi100623g.
10
Structural analysis and unique molecular recognition properties of a Bauhinia forficata lectin that inhibits cancer cell growth.一种抑制癌细胞生长的羊蹄甲凝集素的结构分析及独特分子识别特性
FEBS J. 2017 Feb;284(3):429-450. doi: 10.1111/febs.13989. Epub 2017 Feb 1.
引用本文的文献
1
Asialoglycoprotein receptor 1: a multifaceted receptor in the liver and cardiovascular system.去唾液酸糖蛋白受体1：肝脏和心血管系统中的多功能受体。
Front Med (Lausanne). 2025 Aug 7;12:1653452. doi: 10.3389/fmed.2025.1653452. eCollection 2025.
2
MOTAI: A Novel Method for the Study of O-GalNAcylation and Complex O-Glycosylation in Cancer.MOTAI：一种研究癌症中 O-GalNAc 糖基化和复杂 O-糖基化的新方法。
Anal Chem. 2024 Jul 16;96(28):11137-11145. doi: 10.1021/acs.analchem.3c05018. Epub 2024 Jul 2.
3
Mass Spectrometry Strategies for -Glycoproteomics.用于糖蛋白质组学的质谱策略
Cells. 2024 Feb 25;13(5):394. doi: 10.3390/cells13050394.
4
Mass spectrometry-based methods for investigating the dynamics and organization of the surfaceome: exploring potential clinical implications.基于质谱的方法研究表面组的动态和组织：探索潜在的临床意义。
Expert Rev Proteomics. 2024 Jan-Mar;21(1-3):99-113. doi: 10.1080/14789450.2024.2314148. Epub 2024 Feb 9.
5
Systematic analysis of the impact of phosphorylation and O-GlcNAcylation on protein subcellular localization.系统分析磷酸化和 O-GlcNAc 化对蛋白质亚细胞定位的影响。
Cell Rep. 2023 Jul 25;42(7):112796. doi: 10.1016/j.celrep.2023.112796. Epub 2023 Jul 14.
6
Deciphering Protein O-GalNAcylation: Method Development and Disease Implication.解析蛋白质O-连接N-乙酰半乳糖胺糖基化：方法开发与疾病关联
ACS Omega. 2023 May 24;8(22):19223-19236. doi: 10.1021/acsomega.3c01653. eCollection 2023 Jun 6.
7
Glycomimetic Peptides as Therapeutic Tools.拟糖肽作为治疗工具。
Pharmaceutics. 2023 Feb 17;15(2):688. doi: 10.3390/pharmaceutics15020688.
8
Recent development of analytical methods for disease-specific protein -GlcNAcylation.疾病特异性蛋白质O-连接N-乙酰葡糖胺化分析方法的最新进展。
RSC Adv. 2022 Dec 21;13(1):264-280. doi: 10.1039/d2ra07184c. eCollection 2022 Dec 19.
9
Recombinant mucin biotechnology and engineering.重组黏蛋白生物技术与工程。
Adv Drug Deliv Rev. 2023 Feb;193:114618. doi: 10.1016/j.addr.2022.114618. Epub 2022 Nov 11.
10
Cancer glycomics offers potential biomarkers and therapeutic targets in the framework of 3P medicine.癌症糖组学为 3P 医学框架下的潜在生物标志物和治疗靶点提供了可能。
Front Endocrinol (Lausanne). 2022 Aug 22;13:970489. doi: 10.3389/fendo.2022.970489. eCollection 2022.
本文引用的文献
1
Quantitative investigation of human cell surface -glycoprotein dynamics.人类细胞表面糖蛋白动力学的定量研究。
Chem Sci. 2017 Jan 1;8(1):268-277. doi: 10.1039/c6sc01814a. Epub 2016 Aug 15.
2
Murine Plasma N-Glycosylation Traits Associated with Sex and Strain.与性别和品系相关的小鼠血浆N-糖基化特征
J Proteome Res. 2016 Oct 7;15(10):3489-3499. doi: 10.1021/acs.jproteome.6b00071. Epub 2016 Sep 2.
3
N-linked glycosite profiling and use of Skyline as a platform for characterization and relative quantification of glycans in differentiating xylem of Populus trichocarpa.N-连接糖基化位点分析及 Skyline 平台在鉴定和相对定量毛白杨木质部聚糖中的应用。
Anal Bioanal Chem. 2017 Jan;409(2):487-497. doi: 10.1007/s00216-016-9776-5. Epub 2016 Aug 4.
4
A Novel Quantitative Mass Spectrometry Platform for Determining Protein O-GlcNAcylation Dynamics.一种用于测定蛋白质O-连接N-乙酰葡糖胺糖基化动力学的新型定量质谱平台。
Mol Cell Proteomics. 2016 Jul;15(7):2462-75. doi: 10.1074/mcp.O115.049627. Epub 2016 Apr 25.
5
Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions Provide Evidence of the Glycan Structure.糖肽产生的鎓离子独特的串联质谱（MS/MS）碎裂途径为聚糖结构提供了证据。
Chemistry. 2016 Jan 18;22(3):1114-24. doi: 10.1002/chem.201503659. Epub 2015 Dec 11.
6
Extended O-GlcNAc on HLA Class-I-Bound Peptides.HLA I类结合肽上的O-连接N-乙酰葡糖胺延伸修饰
J Am Chem Soc. 2015 Sep 2;137(34):10922-10925. doi: 10.1021/jacs.5b06586. Epub 2015 Aug 19.
7
Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides.通过糖肽的液相色谱-串联质谱中氧鎓离子碎裂图谱分析确定糖类结构
J Proteome Res. 2014 Dec 5;13(12):6024-32. doi: 10.1021/pr500898r. Epub 2014 Nov 17.
8
A universal chemical enrichment method for mapping the yeast N-glycoproteome by mass spectrometry (MS).一种通过质谱法（MS）绘制酵母N-糖蛋白质组图谱的通用化学富集方法。
Mol Cell Proteomics. 2014 Jun;13(6):1563-72. doi: 10.1074/mcp.M113.036251. Epub 2014 Apr 1.
9
N-Glycosylation site analysis of proteins from Saccharomyces cerevisiae by using hydrophilic interaction liquid chromatography-based enrichment, parallel deglycosylation, and mass spectrometry.利用亲水作用色谱法基于富集、平行去糖基化和质谱法分析酿酒酵母蛋白质的 N-糖基化位点。
J Proteome Res. 2014 Mar 7;13(3):1485-93. doi: 10.1021/pr401049e. Epub 2014 Feb 21.
10
Glycotherapy: new advances inspire a reemergence of glycans in medicine.糖疗法：新进展激发了聚糖在医学领域的再度兴起。
Chem Biol. 2014 Jan 16;21(1):16-37. doi: 10.1016/j.chembiol.2013.09.010. Epub 2013 Nov 21.
Zotero 插件