Cell-specific bioorthogonal tagging of glycoproteins.
Cell-specific bioorthogonal tagging of glycoproteins.
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DOI：
10.1038/s41467-022-33854-0
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发表时间：
2022-10-25
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
糖蛋白表达的改变是癌症发展无可争议的必然结果。理解这些变化是至关重要的，但受到细胞模型系统的限制。例如，肿瘤和宿主之间复杂的相互作用不能在肿瘤衍生细胞系的单一培养中充分再现。更复杂的共培养模型通常依赖于蛋白质组分析的分选程序，很少捕获蛋白质糖基化的细节。在这里，我们报告了一种称为生物正交细胞系特异性糖蛋白标记（BOCTAG）的策略。通过用人工生物合成途径转染来装备细胞，所述人工生物合成途径将生物正交标记的糖转化成相应的核苷酸-糖。只有转染的细胞在非转染细胞存在下将生物正交标签掺入糖蛋白中。我们采用BOCTAG作为一种成像技术，并在质谱-糖蛋白组学中注释细胞特异性糖基化位点。我们展示了在共培养和小鼠模型中的应用，允许分析糖蛋白组作为细胞功能的重要调节剂。糖蛋白表达的变化与疾病相关，但分泌的糖蛋白不能准确地追溯到它们的细胞系来源。在这里，作者开发了一种策略，在共培养系统和体内以细胞系特异性方式化学标记和分析糖蛋白。
Altered glycoprotein expression is an undisputed corollary of cancer development. Understanding these alterations is paramount but hampered by limitations underlying cellular model systems. For instance, the intricate interactions between tumour and host cannot be adequately recapitulated in monoculture of tumour-derived cell lines. More complex co-culture models usually rely on sorting procedures for proteome analyses and rarely capture the details of protein glycosylation. Here, we report a strategy termed Bio-Orthogonal Cell line-specific Tagging of Glycoproteins (BOCTAG). Cells are equipped by transfection with an artificial biosynthetic pathway that transforms bioorthogonally tagged sugars into the corresponding nucleotide-sugars. Only transfected cells incorporate bioorthogonal tags into glycoproteins in the presence of non-transfected cells. We employ BOCTAG as an imaging technique and to annotate cell-specific glycosylation sites in mass spectrometry-glycoproteomics. We demonstrate application in co-culture and mouse models, allowing for profiling of the glycoproteome as an important modulator of cellular function. Changes in glycoprotein expression are correlates of disease, but secreted glycoproteins cannot be accurately traced to their cell line of origin. Here, the authors develop a strategy to chemically tag and profile glycoproteins in a cell line-specific manner in co-culture systems and in vivo.
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影响因子：
4.3
作者：
Kailemia, Muchena J.;Park, Dayoung;Lebrilla, Carlito B.
通讯作者：
Lebrilla, Carlito B.
影响因子：
48
作者：
Gauthier, Nicholas P.;Soufi, Boumediene;Walkowicz, William E.;Pedicord, Virginia A.;Mavrakis, Konstantinos J.;Macek, Boris;Gin, David Y.;Sander, Chris;Millerephrin, Martin L.
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Millerephrin, Martin L.
影响因子：
64.5
作者：
KINGSLEY, DM;KOZARSKY, KF;KRIEGER, M
通讯作者：
KRIEGER, M
影响因子：
4
作者：
Lewis YE;Galesic A;Levine PM;De Leon CA;Lamiri N;Brennan CK;Pratt MR
通讯作者：
Pratt MR
影响因子：
7.8
作者：
Cioce A;Malaker SA;Schumann B
通讯作者：
Schumann B