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Using terahertz spectroscopy as a protein binding assay

机译:使用太赫兹光谱作为蛋白质结合测定

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摘要

The vibrational modes corresponding to protein tertiary structural motion lay in the far infrared or terahertz frequency range. These collective large scale motions depend on global structure and thus will necessarily be perturbed by ligand binding events. We discuss the use of terahertz dielectric spectroscopy to measure these vibrational modes and the sensitivity of the technique to changes in protein conformation, oxidation state and environment. A challenge of applying this sensitivity as a spectroscopic assay for ligand binding is the sensitivity of the technique to both bulk water and water bound to the protein. This sensitivity can entirely obscure the signal from the protein or protein-ligand complex itself, thus necessitating sophisticated sample preparation making the technique impractical for industrial applications. We discuss methods to overcome this background and demonstrate how terahertz spectroscopy can be used to quickly assay protein binding for proteomics and pharmaceutical research.
机译:与蛋白质三级结构运动相对应的振动模式位于远红外或太赫兹频率范围内。这些集体的大规模运动取决于整体结构,因此必然会受到配体结合事件的干扰。我们讨论了使用太赫兹介电谱法测量这些振动模式以及该技术对蛋白质构象,氧化态和环境变化的敏感性。将这种敏感性用作用于配体结合的光谱测定法的挑战是该技术对大量水和与蛋白质结合的水的敏感性。这种敏感性可以完全掩盖来自蛋白质或蛋白质-配体复合物本身的信号,因此需要进行复杂的样品制备,从而使该技术不适用于工业应用。我们讨论了克服这一背景的方法,并证明了太赫兹光谱技术可用于快速分析蛋白质结合的蛋白质组学和药物研究。

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