High-throughput site-specific N-glycoproteomics reveals glyco-signatures for liver disease diagnosis.

High-throughput site-specific N-glycoproteomics reveals glyco-signatures for liver disease diagnosis.
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DOI:
10.1093/nsr/nwac059
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发表时间:
2023-01
影响因子:
20.6
通讯作者:
--
中科院分区:
综合性期刊1区
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--
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糖蛋白组已经成为筛选生物标记物的重要靶点,因为糖基化改变是癌细胞的一个标志。在这项工作中,我们将串联质量标记结合到定量糖蛋白组学中,开发了一种化学标记辅助的互补解离方法,用于完整N-糖肽的多重分析。利用两种不同的鉴定方法和多重标记方法的互补性,我们对人血清免疫球蛋白G(Ig G)进行了迄今为止最全面的位点特异性和亚类特异性N糖基化分析。通过对90例不同严重程度的肝病患者和健康人血清的分析,发现IgG1-H3N5F1和IgG4-H4N3联合检测可用于肝病不同阶段的鉴别诊断。最后,我们使用有针对性的平行反应监测,成功地验证了糖基化在肝病中的表达变化在包括45个血清样本的不同样本队列中。建立了一种高通量完整糖肽定量策略(HTiGQs)。HTiGQs从免疫球蛋白中发现了几个糖基信号,可用于诊断和区分肝病的不同阶段。
The glycoproteome has emerged as a prominent target for screening biomarkers, as altered glycosylation is a hallmark of cancer cells. In this work, we incorporated tandem mass tag labeling into quantitative glycoproteomics by developing a chemical labeling-assisted complementary dissociation method for the multiplexed analysis of intact N-glycopeptides. Benefiting from the complementary nature of two different mass spectrometry dissociation methods for identification and multiplex labeling for quantification of intact N-glycopeptides, we conducted the most comprehensive site-specific and subclass-specific N-glycosylation profiling of human serum immunoglobulin G (IgG) to date. By analysing the serum of 90 human patients with varying severities of liver diseases, as well as healthy controls, we identified that the combination of IgG1-H3N5F1 and IgG4-H4N3 can be used for distinguishing between different stages of liver diseases. Finally, we used targeted parallel reaction monitoring to successfully validate the expression changes of glycosylation in liver diseases in a different sample cohort that included 45 serum samples. A high-throughput intact glycopeptide quantification strategy (HTiGQs) is developed. HTiGQs identified several glyco-signatures from IgG that can be used for diagosis and distinguishing different stages of liver diseases.
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期刊: Molecular & cellular proteomics : MCP
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