Benefits of Chemical Sugar Modifications Introduced by Click Chemistry for Glycoproteomic Analyses.
Benefits of Chemical Sugar Modifications Introduced by Click Chemistry for Glycoproteomic Analyses.
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DOI：
10.1021/jasms.1c00084
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发表时间：
2021-09-01
影响因子：
3.2
通讯作者：
Malaker SA
中科院分区：
文献类型：
--
作者：
Calle B;Bineva-Todd G;Marchesi A;Flynn H;Ghirardello M;Tastan OY;Roustan C;Choi J;Galan MC;Schumann B;Malaker SA
粘蛋白O-糖基化是最复杂的翻译后修饰之一。尽管参与了许多生理过程，但与其他修饰相比，O-糖基化仍然没有得到充分的研究，这仅仅是因为缺乏正确的分析工具。特别是，由于O-糖基化缺乏一致的基序，糖肽的电荷密度较低，从而影响了ETD的断裂，以及修饰多肽的糖链结构不可预测等原因，用质谱法分析完整的O-糖肽是具有挑战性的。最近，我们引入了化学修饰的单糖类似物，允许在生物素基浓缩手柄进行生物正交衍生化后选择性地跟踪和表征粘蛋白O-葡聚糖。在这样做的过程中，我们意识到在这些研究中使用的化学修饰具有额外的好处，允许改进串联质谱仪的分析。在这项工作中，我们通过产生一系列新的GalNAc类似糖肽来建立这一发现。我们表征了这些修饰糖肽的质谱学特征及其生物正交富集试剂留下的特征残基。我们的数据表明，糖肽谱的化学方法提供了优化属性的机会，例如增加的电荷状态、更高的电荷密度和可预测的碎裂行为。
Mucin-type O-glycosylation is among the most complex post-translational modifications. Despite mediating many physiological processes, O-glycosylation remains understudied compared to other modifications, simply because the right analytical tools are lacking. In particular, analysis of intact O-glycopeptides by mass spectrometry is challenging for several reasons; O-glycosylation lacks a consensus motif, glycopeptides have low charge density which impairs ETD fragmentation, and the glycan structures modifying the peptides are unpredictable. Recently, we introduced chemically modified monosaccharide analogs that allowed selective tracking and characterization of mucin-type O-glycans after bioorthogonal derivatization with biotin-based enrichment handles. In doing so, we realized that the chemical modifications used in these studies have additional benefits that allow for improved analysis by tandem mass spectrometry. In this work, we built on this discovery by generating a series of new GalNAc analog glycopeptides. We characterized the mass spectrometric signatures of these modified glycopeptides and their signature residues left by bioorthogonal enrichment reagents. Our data indicate that chemical methods for glycopeptide profiling offer opportunities to optimize attributes such as increased charge state, higher charge density, and predictable fragmentation behavior.
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DOI：
10.1074/mcp.r120.002277
发表时间：
2021
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Riley NM;Bertozzi CR;Pitteri SJ
通讯作者：
Pitteri SJ
影响因子：
10.1
作者：
Malaker SA;Penny SA;Steadman LG;Myers PT;Loke JC;Raghavan M;Bai DL;Shabanowitz J;Hunt DF;Cobbold M
通讯作者：
Cobbold M
影响因子：
4.4
作者：
Riley, Nicholas M.;Malaker, Stacy A.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.
影响因子：
4.9
作者：
Galan, M. Carmen;Tran, Anh Tuan;Bernard, Claire
通讯作者：
Bernard, Claire
影响因子：
7.8
作者：
Cioce A;Malaker SA;Schumann B
通讯作者：
Schumann B