Dissecting Multivalent Lectin-Carbohydrate Recognition Using Polyvalent Multifunctional Glycan-Quantum Dots.
Dissecting Multivalent Lectin-Carbohydrate Recognition Using Polyvalent Multifunctional Glycan-Quantum Dots.
复制标题
DOI：
10.1021/jacs.7b05104
复制
发表时间：
2017-08-30
影响因子：
15
通讯作者：
Zhou D
中科院分区：
文献类型：
--
作者：
Guo Y;Nehlmeier I;Poole E;Sakonsinsiri C;Hondow N;Brown A;Li Q;Li S;Whitworth J;Li Z;Yu A;Brydson R;Turnbull WB;Pöhlmann S;Zhou D
多价蛋白质-碳水化合物相互作用启动病毒/细菌与靶细胞之间的第一次接触，最终导致感染。了解所涉及的结构和结合模式对于设计特异性、有效的多价抑制剂至关重要。然而，缺乏关于这种灵活、复杂和多聚体细胞表面膜蛋白的结构信息常常阻碍了此类努力。在此，我们报告，量子点（QDs）显示与密集阵列的单糖/二糖是强大的探针多价蛋白质-聚糖相互作用。使用一对密切相关的四聚体凝集素，DC-SIGN和DC-SIGNR，它们与HIV和埃博拉病毒糖蛋白（EBOV-GP）结合以增强病毒进入并感染靶细胞，我们表明这种量子点有效地剖析了不同的DC-SIGN/R-聚糖结合模式（四-/二-/单价）通过多模式读出的组合：Förster共振能量转移（FRET），流体动力学尺寸测量和透射电子显微镜成像。我们还报告了一种新的QD-FRET方法，用于定量QD-DC-SIGN/R结合亲和力，揭示DC-SIGN与QD的结合比DC-SIGNR紧密100倍。这一结果与DC-SIGN对某些HIV毒株的转感染效率高于DC-SIGNR相一致。最后，我们表明QD有效地抑制DC-SIGN介导的EBOV-GP驱动的靶细胞转导的增强，IC 50值低至0.7 nM，与通过FRET测量的其DC-SIGN结合常数（表观Kd = 0.6 nM）很好地匹配。这些结果表明，聚糖量子点是强大的多功能探针，用于解剖多价蛋白质-配体识别和预测在细胞水平上的病毒感染的糖纳米颗粒抑制。
Multivalent protein–carbohydrate interactions initiate the first contacts between virus/bacteria and target cells, which ultimately lead to infection. Understanding the structures and binding modes involved is vital to the design of specific, potent multivalent inhibitors. However, the lack of structural information on such flexible, complex, and multimeric cell surface membrane proteins has often hampered such endeavors. Herein, we report that quantum dots (QDs) displayed with a dense array of mono-/disaccharides are powerful probes for multivalent protein–glycan interactions. Using a pair of closely related tetrameric lectins, DC-SIGN and DC-SIGNR, which bind to the HIV and Ebola virus glycoproteins (EBOV-GP) to augment viral entry and infect target cells, we show that such QDs efficiently dissect the different DC-SIGN/R-glycan binding modes (tetra-/di-/monovalent) through a combination of multimodal readouts: Förster resonance energy transfer (FRET), hydrodynamic size measurement, and transmission electron microscopy imaging. We also report a new QD-FRET method for quantifying QD-DC-SIGN/R binding affinity, revealing that DC-SIGN binds to the QD >100-fold tighter than does DC-SIGNR. This result is consistent with DC-SIGN’s higher trans-infection efficiency of some HIV strains over DC-SIGNR. Finally, we show that the QDs potently inhibit DC-SIGN-mediated enhancement of EBOV-GP-driven transduction of target cells with IC50 values down to 0.7 nM, matching well to their DC-SIGN binding constant (apparent Kd = 0.6 nM) measured by FRET. These results suggest that the glycan-QDs are powerful multifunctional probes for dissecting multivalent protein–ligand recognition and predicting glyconanoparticle inhibition of virus infection at the cellular level.
登录
查看更多内容
影响因子：
46.2
作者：
Kiessling LL;Grim JC
通讯作者：
Grim JC
影响因子：
64.5
作者：
Geijtenbeek, TBH;Torensma, R;Figdor, CG
通讯作者：
Figdor, CG
影响因子：
16.6
作者：
Branson, Thomas R.;McAllister, Tom E.;Garcia-Hartjes, Jaime;Fascione, Martin A.;Ross, James F.;Warriner, Stuart L.;Wennekes, Tom;Zuilhof, Han;Turnbull, W. Bruce
通讯作者：
Turnbull, W. Bruce
影响因子：
15
作者：
Bhatia, Sumati;Camacho, Luis Cuellar;Haag, Rainer
通讯作者：
Haag, Rainer
影响因子：
2.9
作者：
Kong, Leopold;Wilson, Ian A.;Kwong, Peter D.
通讯作者：
Kwong, Peter D.