Grayfer Tatyana D, Yamani Khalil, Jung Erik, Chesnokov Gleb A, Ferrara Isabella, Hsiao Chien-Chi, Georgiou Antri, Michel Jeremy, Bailly Aurélien, Sieber Simon, Eberl Leo, Gademann Karl
Department of Chemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland.
Department of Plant and Microbial Biology, University of Zurich, Zollikerstrasse 107, 8008 Zürich, Switzerland.
JACS Au. 2023 Mar 20;3(4):1151-1161. doi: 10.1021/jacsau.3c00037. eCollection 2023 Apr 24.
Allylic cyclitols were investigated as covalent inhibitors of glycoside hydrolases by chemical, enzymatic, proteomic, and computational methods. This approach was inspired by the C cyclitol natural product streptol glucoside, which features a potential carbohydrate leaving group in the 4-position (carbohydrate numbering). To test this hypothesis, carbocyclic inhibitors with leaving groups in the 4- and 6- positions were prepared. The results of enzyme kinetics analyses demonstrated that dinitrophenyl ethers covalently inhibit α-glucosidases of the GH13 family without reactivation. The labeled enzyme was studied by proteomics, and the active site residue Asp214 was identified as modified. Additionally, computational studies, including enzyme homology modeling and density functional theory (DFT) calculations, further delineate the electronic and structural requirements for activity. This study demonstrates that previously unexplored 4- and 6-positions can be exploited for successful inhibitor design.
通过化学、酶学、蛋白质组学和计算方法，对烯丙基环糖醇作为糖苷水解酶的共价抑制剂进行了研究。这种方法的灵感来源于C环糖醇天然产物链霉葡萄糖苷，其在4位（碳水化合物编号）具有潜在的碳水化合物离去基团。为了验证这一假设，制备了在4位和6位带有离去基团的碳环抑制剂。酶动力学分析结果表明，二硝基苯基醚可共价抑制GH13家族的α-葡萄糖苷酶，且不会再活化。通过蛋白质组学研究了标记的酶，并确定活性位点残基Asp214被修饰。此外，包括酶同源建模和密度泛函理论（DFT）计算在内的计算研究，进一步阐明了活性的电子和结构要求。这项研究表明，以前未被探索的4位和6位可用于成功的抑制剂设计。
