Biomedical sciences Division, School of Medicine, University of California, Riverside, 900 University Avenue, CA, 92521, Riverside, USA.
ChemMedChem. 2020 Nov 18;15(22):2176-2184. doi: 10.1002/cmdc.202000355. Epub 2020 Oct 13.
Recently we reported on aryl-fluorosulfates as possible stable and effective electrophiles for the design of lysine covalent, cell permeable antagonists of protein-protein interactions (PPIs). Here we revisit the use of aryl-sulfonyl fluorides as Lys-targeting moieties, incorporating these electrophiles in XIAP (X-linked inhibitor of apoptosis protein) targeting agents. We evaluated stability in buffer and reactivity with Lys311 of XIAP of various aryl-sulfonyl fluorides using biochemical and biophysical approaches, including displacement assays, mass spectrometry, SDS gel electrophoresis, and denaturation thermal shift measurements. To assess whether these modified electrophilic "warheads" can also react with Tyr, we repeated these evaluations with a Lys311Tyr XIAP mutant. Using a direct cellular assay, we could demonstrate that selected agents are cell permeable and interact covalently with their intended target in cell. These results suggest that certain substituted aryl-sulfonyl fluorides can be useful Lys- or Tyr-targeting electrophiles for the design of covalent pharmacological tools or even future therapeutics targeting protein-protein interactions.
最近，我们报道了芳基氟硫酸盐作为设计赖氨酸共价、细胞渗透的蛋白质-蛋白质相互作用（PPIs）拮抗剂的可能稳定且有效的亲电试剂。在这里，我们重新探讨了芳基磺酰氟作为赖氨酸靶向部分的用途，将这些亲电试剂纳入 XIAP（凋亡蛋白抑制因子 X 连锁）靶向剂中。我们使用生化和生物物理方法，包括取代测定、质谱、SDS 凝胶电泳和变性热移测量，评估了各种芳基磺酰氟在缓冲液中的稳定性以及与 XIAP 的赖氨酸 311 的反应性。为了评估这些修饰的亲电“弹头”是否也可以与 Tyr 反应，我们用 Lys311Tyr XIAP 突变体重复了这些评估。使用直接细胞测定，我们可以证明选定的试剂是细胞渗透的，并且在细胞中与它们的预期靶标发生共价相互作用。这些结果表明，某些取代的芳基磺酰氟可以作为赖氨酸或 Tyr 靶向亲电试剂，用于设计共价药理学工具，甚至是针对蛋白质-蛋白质相互作用的未来治疗药物。
