Solution and crystallographic studies of branched multivalent ligands that inhibit the receptor-binding of cholera toxin.
Solution and crystallographic studies of branched multivalent ligands that inhibit the receptor-binding of cholera toxin.
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DOI：
10.1021/ja027584k
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发表时间：
2002-10
影响因子：
15
通讯作者：
Zhongsheng Zhang;E. Merritt;M. Ahn;C. Roach;Z. Hou;C. Verlinde;W. Hol;E. Fan
中科院分区：
文献类型：
--
作者：
Zhongsheng Zhang;E. Merritt;M. Ahn;C. Roach;Z. Hou;C. Verlinde;W. Hol;E. Fan
基于结构的多价配体设计为高亲和力蛋白抑制剂提供了一种有吸引力的策略。霍乱毒素是严重腹泻病霍乱的病原体，也是 AB(5) 细菌毒素家族的成员，其受体结合位点的空间排列为设计具有 5 重对称性的分支多价配体提供了机会。我们的模块化合成能够构建一系列复杂的配体，该配体具有五个柔性臂，每个臂以二价配体结尾。这些配体中最大的分子量为 10.6 kDa。这些配体能够以高亲和力同时结合两个毒素B五聚体分子，从而阻断霍乱毒素的受体结合过程。使用最好的支链十价配体，其抑制能力比单价配体提高了数百万倍以上。这比之前观察到的相应非支链五价配体的改进要好。动态光散射研究表明，溶液中会形成浓度依赖性的独特 1:1 和 1:2 配体/毒素复合物，且没有非特异性聚集的迹象。这与以 1.45 A 分辨率解析的支链多价配体/毒素 B 五聚体复合物的晶体结构完全一致，该结构显示了固态下特定的 1:2 配体/毒素复合物的形成。这些结果重申了基于结构的多价蛋白质配体设计作为实现高亲和力和有效抑制的一般策略的力量。
The structure-based design of multivalent ligands offers an attractive strategy toward high affinity protein inhibitors. The spatial arrangement of the receptor-binding sites of cholera toxin, the causative agent of the severe diarrheal disease cholera and a member of the AB(5) bacterial toxin family, provides the opportunity of designing branched multivalent ligands with 5-fold symmetry. Our modular synthesis enabled the construction of a family of complex ligands with five flexible arms each ending with a bivalent ligand. The largest of these ligands has a molecular weight of 10.6 kDa. These ligands are capable of simultaneously binding to two toxin B pentamer molecules with high affinity, thus blocking the receptor-binding process of cholera toxin. A more than million-fold improvement over the monovalent ligand in inhibitory power was achieved with the best branched decavalent ligand. This is better than the improvement observed earlier for the corresponding nonbranched pentavalent ligand. Dynamic light scattering studies demonstrate the formation of concentration-dependent unique 1:1 and 1:2 ligand/toxin complexes in solution with no sign of nonspecific aggregation. This is in complete agreement with a crystal structure of the branched multivalent ligand/toxin B pentamer complex solved at 1.45 A resolution that shows the specific 1:2 ligand/toxin complex formation in the solid state. These results reiterate the power of the structure-based design of multivalent protein ligands as a general strategy for achieving high affinity and potent inhibition.