Using Biotinylated myo-Inositol Hexakisphosphate to Investigate Inositol Pyrophosphate–Protein Interactions with Surface-Based Biosensors,Biochemistry

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Using Biotinylated myo-Inositol Hexakisphosphate to Investigate Inositol Pyrophosphate–Protein Interactions with Surface-Based Biosensors
Biochemistry ( IF 2.9 ) Pub Date : 2021-09-09 , DOI: 10.1021/acs.biochem.1c00497
Daniel Couto ₁ , Annika Richter _{2,

3} , Henriette Walter _{2,

3} , David Furkert _{2,

3} , Michael Hothorn ₁ , Dorothea Fiedler _{2,

3}

Affiliation

Inositol pyrophosphates (PP-InsPs) are highly phosphorylated molecules that have emerged as central nutrient messengers in eukaryotic organisms. They can bind to structurally diverse target proteins to regulate biological functions, such as protein–protein interactions. PP-InsPs are strongly negatively charged and interact with highly basic surface patches in proteins, making their quantitative biochemical analysis challenging. Here, we present the synthesis of biotinylated myo-inositol hexakisphosphates and their application in surface plasmon resonance and grating-coupled interferometry assays, to enable the rapid identification, validation, and kinetic characterization of InsP– and PP-InsP–protein interactions.

中文翻译：

使用生物素化的肌醇六磷酸研究肌醇焦磷酸-蛋白质与基于表面的生物传感器的相互作用

肌醇焦磷酸 (PP-InsPs) 是高度磷酸化的分子，已成为真核生物的中枢营养信使。它们可以与结构多样的靶蛋白结合以调节生物功能，例如蛋白质-蛋白质相互作用。PP-InsPs 带强负电荷并与蛋白质中的强碱性表面斑块相互作用，使其定量生化分析具有挑战性。这里，我们提出的生物素化的合成肌肉肌醇hexakisphosphates以及它们在表面等离子体共振和光栅耦合干涉测定法的应用，以使快速识别，验证和InsP-和PP-INSP -蛋白质相互作用的动力学鉴定。

更新日期：2021-09-21

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