Carbohydrate-protein interactions by "clicked" carbohydrate self-assembled monolayers
Carbohydrate-protein interactions by "clicked" carbohydrate self-assembled monolayers
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DOI：
10.1021/ac051919
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发表时间：
2006-03-15
影响因子：
7.4
通讯作者：
Wang, PG
中科院分区：
文献类型：
--
作者：
Zhang, Y;Luo, SZ;Wang, PG
采用Huisgen 1,3-偶极环加成“点击化学”固定化偶氮糖（甘露糖、乳糖、α -半乳糖）在金表面制备碳水化合物自组装单层（sam）。Ibis的制备是基于预先形成的带有炔端基的SAM模板，它可以进一步锚定叠氮糖，形成良好的、稳定的、刚性的糖SAM。点击甘露糖、乳糖和A - gal三糖sam用于分析特定的碳水化合物-蛋白质相互作用（即甘露糖- con A； ecl -乳糖，α - galanti - gal）。经QCM和SPR测定，Con A与甘露糖结合的表观亲和常数分别为（8.7 +/- 2.8）× 10（-5）和（3.9 +/- 0.2）× 10(6) M-1。经QCM测定，乳糖与ECL结合的表观亲和常数为（4.6 +/- 2.4）× 10(6)， a-Gal与多克隆抗gal抗体结合的表观亲和常数为（6.7 +/- 3.3）× 10(6) M-1。SPR、QCM、AFM和电化学研究证实，碳水化合物SAM传感器对其相应的凝集素保持特异性，在点击碳水化合物表面的非特异性吸附可以忽略不计。该研究表明，点击碳水化合物sam与非标记QCM或SPR一起为碳水化合物-蛋白质相互作用的高通量表征提供了一个有效的平台。这样的结合应该是ITC和ELISA等其他方法的有益补充，并为相应的复杂糖生物学过程提供有见地的知识。
A Huisgen 1,3-dipolar cycloaddition "click chemistry" was employed to immobilize azido sugars (mannose, lactose, a-Gal) to fabricate carbohydrate self-assembled monolayers (SAMs) on gold. Ibis fabrication was based on preformed SAM templates incorporated with alkyne terminal groups, which could further anchor the azido sugars to form well-packed, stable, and rigid sugar SAMs. The clicked mannose, lactose, and a-Gal trisaccharide SAMs were used in the analysis of specific carbohydrate-protein interactions (i.e., mannose-Con A; ECL-lactose, alpha-Galanti-Gal). The apparent affinity constant of Con A binding to mannose was (8.7 +/- 2.8) x 10(-5) and (3.9 +/- 0.2) x 10(6) M-1 measured by QCM and SPR, respectively. The apparent affinity constants of lactose binding with ECL and a-Gal binding with polyclonal anti-Gal antibody were determined to be (4.6 +/- 2.4) x 10(6) and (6.7 +/- 3.3) x 10(6) M-1, respectively by QCM. SPR, QCM, AFM, and electrochemistry studies confirmed that the carbohydrate SAM sensors maintained the specificity to their corresponding lectins and nonspecific adsorption on the clicked carbohydrate surface was negligible. This study showed that the clicked carbohydrate SAMs in concert with nonlabel QCM or SPR offered a potent platform for high-throughput characterization of carbohydrate- protein interactions. Such a combination should complement other methods such as ITC and ELISA in a favorable manner and provide insightful knowledge for the corresponding complex glycobiological processes.