Thiophosphate bioisosteres of inositol hexakisphosphate enhance binding affinity and residence time on bacterial virulence factors†

IF 3.1 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Rebecca Cummer, Garvit Bhatt, Lauren M. Finn, Bettina G. Keller, Bhushan Nagar and Bastien Castagner
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Abstract

Inositol phosphates are essential for mammalian cell signalling with critical roles in cellular processes. The fully phosphorylated inositol phosphate, myo-inositol hexakisphosphate (IP6), modulates numerous eukaryotic proteins and bacterial virulence factors. It has been suggested that the high charge density of IP6 causes restructuring of virulence factors in mammalian cells, activating their enzymatic activity. IP6 is challenging to study due to its phytase instability and propensity to precipitate. Here we suggest that the thiophosphate bioisostere, myo-inositol hexakisthiophosphate (IT6), will mitigate these issues, as thiophosphate substitution has been found to be phytase resistant and improve solubility. Assessment of the chemical properties of IT6 has indeed validated these characteristics. In addition, we performed biophysical characterization of IT6 binding to the virulence factors Salmonella enterica serovar Typhimurium AvrA, Vibrio parahaemolyticus VopA, and Clostridioides difficile TcdB. Our data show that the higher charge density of IT6 increased its binding affinity and residence time on the proteins, which improved stabilization of the bound-state. IT6 is a valuable tool for structural biology research and the described biophysical characteristics of thiophosphate substitution are of value in medicinal chemistry.

Abstract Image

六磷酸肌醇的硫代磷酸生物异构体增强了对细菌毒力因子的结合亲和力和停留时间。
肌醇磷酸酯是哺乳动物细胞信号传递所必需的,在细胞过程中起着关键作用。完全磷酸化的磷酸肌醇,肌六磷酸肌醇(IP6),调节许多真核蛋白和细菌毒力因子。有研究认为,IP6的高电荷密度导致哺乳动物细胞中毒力因子的重组,激活它们的酶活性。IP6由于其植酸酶不稳定性和沉淀倾向而具有挑战性。在这里,我们建议硫代磷酸生物异构体,肌肌醇己基硫代磷酸(IT6),将缓解这些问题,因为硫代磷酸替代已被发现具有抗植酸酶和改善溶解度的能力。对IT6化学性质的评估确实证实了这些特性。此外,我们还对IT6与肠沙门氏菌血清鼠伤寒沙门氏菌AvrA、副溶血性弧菌VopA和艰难梭菌TcdB等毒力因子的结合进行了生物物理表征。我们的数据表明,较高的电荷密度增加了IT6在蛋白质上的结合亲和力和停留时间,从而提高了结合态的稳定性。IT6是结构生物学研究的一个有价值的工具,所描述的硫代磷酸盐取代的生物物理特性在药物化学中具有价值。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
6.10
自引率
0.00%
发文量
128
审稿时长
10 weeks
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