Introduction of C-alkyl branches to L-iminosugars changes their active site binding orientation
Introduction of C-alkyl branches to L-iminosugars changes their active site binding orientation
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将 C-烷基分支引入 L-亚氨基糖改变了它们的活性位点结合方向
DOI：
10.1039/d2ob01099b
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发表时间：
2022
影响因子：
3.2
通讯作者：
Yu Chu-Yi
中科院分区：
文献类型：
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作者：
Kato Atsushi;Nakagome Izumi;Yoshimura Kosuke;Kanekiyo Uta;Kishida Mana;Shinzawa Kenta;Lu Tian-Tian;Li Yi-Xian;Nash Robert J.;Fleet George W. J.;Tanaka Nobutada;Yu Chu-Yi
L-ido-Deoxynojirimycin (L-ido-DNJ) 本身对人溶酶体酸性 α-葡萄糖苷酶 (GAA) 没有亲和力，而 5-C-甲基-L-ido-DNJ 对 GAA 有很强的亲和力，与葡萄糖类似物 DNJ 相当，Ki 值为 0.060 μM。仅当引入甲基和乙基时，才能观察到这种对 GAA 的优异亲和力和酶稳定性。对接模拟分析表明，5-C-烷基-L-ido-DNJs的烷基链根据其长度存储在三个不同的口袋中，从而改变了分子取向。 DNJ和5-C-甲基-L-ido-DNJ的结合位姿比较表明，它们与Asp404、Asp518和Asp616形成共同的离子相互作用，但结合方向和配体与每个氨基酸残基之间的距离不同。 5-C-Mmethyl-L-ido-DNJ 剂量依赖性地增加具有 M519V 突变的 Pompe 患者成纤维细胞的细胞内 GAA 活性，并促进酶转运至溶酶体。这项研究提供了通过将烷基分支引入稀有糖和L-糖型亚氨基糖来改变结合方向来设计高亲和力配体的策略的第一个例子。
L-ido-Deoxynojirimycin (L-ido-DNJ) itself showed no affinity for human lysosomal acid α-glucosidase (GAA), whereas 5-C-methyl-L-ido-DNJ showed a strong affinity for GAA, comparable to the glucose analog DNJ, with a Ki value of 0.060 μM. This excellent affinity for GAA and enzyme stabilization was observed only when methyl and ethyl groups were introduced. Docking simulation analysis revealed that the alkyl chains of 5-C-alkyl-L-ido-DNJs were stored in three different pockets, depending on their length, thereby the molecular orientation was changed. Comparison of the binding poses of DNJ and 5-C-methyl-L-ido-DNJ showed that they formed a common ionic interaction with Asp404, Asp518, and Asp616, but both the binding orientation and the distance between the ligand and each amino acid residue were different. 5-C-Methyl-L-ido-DNJ dose-dependently increased intracellular GAA activity in Pompe patient fibroblasts with the M519V mutation and also promoted enzyme transport to lysosomes. This study provides the first example of a strategy to design high-affinity ligands by introducing alkyl branches into rare sugars and L-sugar-type iminosugars to change the orientation of binding.