A photoaffinity glycan-labeling approach to investigate immunoglobulin glycan-binding partners.
A photoaffinity glycan-labeling approach to investigate immunoglobulin glycan-binding partners.
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DOI：
10.1093/glycob/cwad055
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发表时间：
2023-10-29
期刊：
影响因子：
4.3
通讯作者：
中科院分区：
文献类型：
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聚糖在生物学中发挥着关键作用。然而，由于聚糖-蛋白质相互作用的亲和力较低，许多相互作用对仍然未知。 B 细胞生物学中涉及的两种重要糖蛋白是 B 细胞受体及其分泌的对应物抗体。研究表明，这些 B 细胞特异性分子表达的聚糖可以通过聚糖结合蛋白调节免疫激活。在多种自身免疫性疾病中，已观察到 IgG 自身抗体可变结构域糖基化的患病率增加。特别是，类风湿性关节炎的标志性自身抗体，即抗瓜氨酸蛋白抗体，携带大量的可变结构域聚糖。这些自身抗体表达的可变域聚糖是 N 连接的、复合型的和 α2-6 唾液酸化的，并且假设携带可变域聚糖的 B 细胞受体通过与聚糖结合蛋白的相互作用促进自身反应性 B 细胞的选择。在这里，我们使用抗瓜氨酸蛋白抗体反应作为原型来研究溶液和原位 B 细胞受体-可变域聚糖相互作用物的潜力。我们使用了 SiaDAz，一种可紫外线激活的唾液酸类似物，带有二氮丙啶部分，可以与近端聚糖结合蛋白形成共价键。我们证明，利用寡糖工程，SiaDAz 可以很容易地整合到抗体和 B 细胞受体的可变域聚糖中。我们的数据显示抗体可变域聚糖能够与抑制性受体 CD22 相互作用。有趣的是，虽然我们没有检测到细胞表面的这种相互作用，但我们捕获了 CD79 β 聚糖 -B 细胞受体的相互作用。这些结果显示了结合光亲和标记和寡糖工程用于识别抗体和 B 细胞受体相互作用的效用，并表明可变域聚糖在我们的测试条件下似乎不是凝集素顺式配体。
Glycans play a pivotal role in biology. However, because of the low-affinity of glycan-protein interactions, many interaction pairs remain unknown. Two important glycoproteins involved in B-cell biology are the B-cell receptor and its secreted counterpart, antibodies. It has been indicated that glycans expressed by these B-cell-specific molecules can modulate immune activation via glycan-binding proteins. In several autoimmune diseases, an increased prevalence of variable domain glycosylation of IgG autoantibodies has been observed. Especially, the hallmarking autoantibodies in rheumatoid arthritis, anti-citrullinated protein antibodies, carry a substantial amount of variable domain glycans. The variable domain glycans expressed by these autoantibodies are N-linked, complex-type, and α2–6 sialylated, and B-cell receptors carrying variable domain glycans have been hypothesized to promote selection of autoreactive B cells via interactions with glycan-binding proteins. Here, we use the anti-citrullinated protein antibody response as a prototype to study potential in solution and in situ B-cell receptor–variable domain glycan interactors. We employed SiaDAz, a UV-activatable sialic acid analog carrying a diazirine moiety that can form covalent bonds with proximal glycan-binding proteins. We show, using oligosaccharide engineering, that SiaDAz can be readily incorporated into variable domain glycans of both antibodies and B-cell receptors. Our data show that antibody variable domain glycans are able to interact with inhibitory receptor, CD22. Interestingly, although we did not detect this interaction on the cell surface, we captured CD79 β glycan–B-cell receptor interactions. These results show the utility of combining photoaffinity labeling and oligosaccharide engineering for identifying antibody and B-cell receptor interactions and indicate that variable domain glycans appear not to be lectin cis ligands in our tested conditions.
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DOI：
10.1038/s41584-022-00867-4
发表时间：
2023-01
期刊：
Nature reviews. Rheumatology
影响因子：
--
作者：
通讯作者：
--
影响因子：
16.6
作者：
Giovannone N;Liang J;Antonopoulos A;Geddes Sweeney J;King SL;Pochebit SM;Bhattacharyya N;Lee GS;Dell A;Widlund HR;Haslam SM;Dimitroff CJ
通讯作者：
Dimitroff CJ
影响因子：
14.2
作者：
Koers, Jana;Sciarrillo, Rocco;Rispens, Theo
通讯作者：
Rispens, Theo
影响因子：
56.9
作者：
Ma, Xinyu;Zhu, Yuwei;Huang, Zhiwei
通讯作者：
Huang, Zhiwei
影响因子：
3.7
作者：
Guhr T;Bloem J;Derksen NI;Wuhrer M;Koenderman AH;Aalberse RC;Rispens T
通讯作者：
Rispens T