Molecular basis of mannose recognition by pradimicins and their application to microbial cell surface imaging
Molecular basis of mannose recognition by pradimicins and their application to microbial cell surface imaging
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普拉霉素识别甘露糖的分子基础及其在微生物细胞表面成像中的应用
DOI：
10.1016/j.chembiol.2019.03.013
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Y.
中科院分区：
文献类型：
--
作者：
Nakagawa;Y.; Doi;T.; Takegoshi;K.; Sugahara;T.; Akase;D.; Aida;M.; Tsuzuki;K.; Watanabe;Y.; Tomura;T.; Ojika;M.; Igarashi;Y.; Hashizume;D.; Ito;Y.
天然产生的帕拉米星(PRMs)对水介质中的d-甘露糖(d-Man)具有特异的识别能力，这是人工小分子所无法实现的。虽然钙离子介导的PRM的二聚化对于它们的d-Man结合是必不可少的，但二聚体的结构还没有被阐明，这使得关于PRM如何识别d-Man的问题仍然悬而未决。因此，我们在这里报告了用X射线结晶学和固体核磁共振光谱相结合的方法来阐明二聚体的结构。再加上我们之前关于PRM的d-Man结合几何的知识，二聚体的阐明使得对d-Man结合模式的可靠估计成为可能。基于该结合模型，我们进一步开发了具有d-Man结合专一性的叠氮功能化PRM衍生物(PRM-叠氮化物)。值得注意的是，PRM-叠氮化物通过与四甲基罗丹明荧光团的结合，使假丝酵母细胞表面含有甘露糖。本研究提供了PRM可作为用于糖生物学研究的凝集素模拟物的实际证明。
Naturally occurring pradimicins (PRMs) show specific recognition of d-mannose (d-Man) in aqueous media, which has never been achieved by artificial small molecules. Although the Ca2+-mediated dimerization of PRMs is essential for their d-Man binding, the dimeric structure has yet to be elucidated, leaving the question open as to how PRMs recognize d-Man. Thus, we herein report the structural elucidation of the dimer by a combination of X-ray crystallography and solid-state NMR spectroscopy. Coupled with our previous knowledge regarding the d-Man binding geometry of PRMs, elucidation of the dimer allowed reliable estimation of the mode of d-Man binding. Based on the binding model, we further developed an azide-functionalized PRM derivative (PRM-Azide) with d-Man binding specificity. Notably, PRM-Azide stainedCandida rugosacells having mannans on their cell surface through conjugation with the tetramethylrhodamine fluorophore. The present study provides the practical demonstration that PRMs can serve as lectin mimics for use in glycobiological studies.