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Ultrasensitive Detection and Characterization of Posttranslational Modifications Using Surface-Enhanced Raman Spectroscopy

机译:使用表面增强拉曼光谱的翻译后修饰的超灵敏检测和表征

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摘要

Posttranslational modification (PTM) of proteins is likely to be the most common mechanism of altering the expression of genetic information. It is essential to characterize PTMs to establish a complete understanding of the activities of proteins. Here, we present a sensitive detection method using surface-enhanced Raman spectroscopy (SERS) that can detect PTMs from as little as zeptomoles of peptide. We demonstrate, using model peptides, the ability of SERS to detect a variety of protein modifications, such as acetylation, trimethylation, phosphorylation, and ubiquitination. In addition, we show the capability to obtain positional information for modifications such as trimethylation and phosphorylation using SERS and wavelet decomposition data analysis techniques. We further show that it is possible to apply SERS to detect PTMs from biological samples such as histones. We envision that this detection method might be a valuable technique that is complementary to mass spectrometry in obtaining orthogonal chemical and modification-specific information from biological samples at sensitive levels.
机译:蛋白质的翻译后修饰(PTM)可能是改变遗传信息表达的最常见机制。表征PTM以建立对蛋白质活性的完整理解至关重要。在这里,我们提出了一种使用表面增强拉曼光谱(SERS)的灵敏检测方法,该方法可以从少至肽的分子中检出PTM。我们证明,使用模型肽,SERS能够检测多种蛋白质修饰,例如乙酰化,三甲基化,磷酸化和泛素化。此外,我们展示了使用SERS和小波分解数据分析技术获得位置信息以进行修饰的能力,例如三甲基化和磷酸化。我们进一步表明,有可能将SERS应用于从生物样品(例如组蛋白)中检测PTM。我们设想,这种检测方法可能是一种有价值的技术,可以与质谱法互补,从而从敏感水平的生物样品中获得正交化学信息和特定于修饰的信息。

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