Dihydroisoxazole Analogs for Labeling and Visualization of Catalytically Active Transglutaminase 2
Dihydroisoxazole Analogs for Labeling and Visualization of Catalytically Active Transglutaminase 2
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DOI：
10.1016/j.chembiol.2010.11.004
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发表时间：
2011-01-28
影响因子：
--
通讯作者：
Khosla, Chaitan
中科院分区：
文献类型：
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作者：
Dafik, Laila;Khosla, Chaitan
我们报告了人转谷氨酰胺酶 2 (TG2)“可点击”抑制剂的合成和初步表征。这些抑制剂具有 3-halo-4,5-二氢异恶唑弹头以及生物正交基团（例如叠氮化物或炔部分），从而能够随后与荧光团进行共价修饰。它们抑制 TG2 的机制基于卤化物置换，从而形成稳定的亚氨基硫醚。针对重组人 TG2 的抑制测定表明，本研究中制备的一些可点击抑制剂具有与先前报道的基准二氢异恶唑抑制剂相当的特异性。在低微摩尔浓度下，它们在 WI-38 成纤维细胞划痕测定中完全抑制瞬时激活的 TG2，随后可用于原位观察活性酶。讨论了这些抑制剂在探讨 TG2 在乳糜泻以及其他疾病中的作用的潜在用途。
We report the synthesis and preliminary characterization of "clickable" inhibitors of human transglutaminase 2 (TG2). These inhibitors possess the 3-halo-4,5-dihydroisoxazole warhead along with bioorthogonal groups such as azide or alkyne moieties that enable subsequent covalent modification with fluorophores. Their mechanism for inhibition of TG2 is based on halide displacement, resulting in the formation of a stable imino thioether. Inhibition assays against recombinant human TG2 revealed that some of the clickable inhibitors prepared in this study have comparable specificity as benchmark dihydroisoxazole inhibitors reported earlier. At low micromolar concentrations they completely inhibited transiently activated TG2 in a WI-38 fibroblast scratch assay and could subsequently be used to visualize the active enzyme in situ. The potential use of these inhibitors to probe the role of TG2 in celiac sprue as well as other diseases is discussed.