Expanding Glycomic Investigations through Thiol-Derivatized Glycans.
Expanding Glycomic Investigations through Thiol-Derivatized Glycans.
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DOI：
10.3390/molecules28041956
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发表时间：
2023-02-18
期刊：
影响因子：
4.6
通讯作者：
Brichacek, Matthew
中科院分区：
文献类型：
--
作者：
Hurst, Robert D. D.;Nieves, Angel;Brichacek, Matthew
关键词：
合成了一种新型聚糖助剂N-（2-硫乙基）-2-氨基苯甲酰胺（TEAB），并对其应用进行了评价。通过水稳定试剂2-吡啶硼烷配合物的还原性胺化反应，将助剂偶联到聚糖上。经反相色谱法纯化，得到的多糖产物为1 ~ 7个连接己糖，收率为60% ~ 90%。这种新型缀合物将一种方便的、货架稳定的硫醇直接引入到所需的游离聚糖上，具有纯化优势，并且可以通过烯烃、卤化物、环氧化物、二硫化物和羧酸盐进行有效的直接修饰，收率为49%至93%。随后，对BSA蛋白进行巯基选择性修饰，生成具有双功能peg -马来酰亚胺连接剂的新糖蛋白。为了进一步说明巯基序的效用，2-硫代吡啶活化含巯基的载体促进了标记聚糖的共价色谱纯化，收率高达63%。最后，通过fitc标记的豆豆蛋白a结合，探索并验证了在光印刷微阵列中实现概念的初步证明。总之，巯基功能化的聚糖极大地扩展了生物偶联工具的多样性，这些工具可以用聚糖开发，并使各种生物学研究成为可能。
N-(2-thioethyl)-2-aminobenzamide (TEAB), a novel glycan auxiliary, was synthesized and its utility was evaluated. The auxiliary was conjugated to glycans by reductive amination with the water-stable reagent 2-picoline borane complex. Glycan products, which ranged from 1 to 7 linked hexoses, were all isolated in yields ranging from 60% to 90% after purification by reverse-phase chromatography. The novel conjugate introduces a convenient, shelf-stable thiol directly onto the desired free glycans with purification advantages and direct modification with efficient reactions through alkenes, halides, epoxides, disulfides, and carboxylates in yields of 49% to 93%. Subsequently, a thiol-selective modification of the BSA protein was used to generate a neoglycoprotein with a bifunctional PEG–maleimide linker. To further illustrate the utility of a thiol motif, 2-thiopyridine activation of a thiol-containing support facilitated the covalent chromatographic purification of labeled glycans in yields up to 63%. Finally, initial proof of concept of implementation in a light printed microarray was explored and validated through FITC-labeled concanavalin A binding. In conclusion, the thiol-functionalized glycans produced greatly expand the diversity of bioconjugation tools that can be developed with glycans and enable a variety of biological investigations.
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影响因子：
16.6
作者：
Shibata K;Natsui S;Tobisu M;Fukumoto Y;Chatani N
通讯作者：
Chatani N
影响因子：
4.4
作者：
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影响因子：
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使用天冬酰胺内肽酶的位点特异性蛋白标记和定义的泛素蛋白结合物的生成。
影响因子：
15
作者：
Fottner, Maximilian;Heimgartner, Johannes;Gantz, Maximilian;Muehlhofer, Rahel;Nast-Kolb, Timon;Lang, Kathrin
通讯作者：
Lang, Kathrin
影响因子：
4.3
作者：
Ruhaak, L. R.;Zauner, G.;Huhn, C.;Bruggink, C.;Deelder, A. M.;Wuhrer, M.
通讯作者：
Wuhrer, M.