Department of Chemistry, University of Massachusetts Amherst, Amherst, Massachusetts 01003, United States.
Molecular & Cellular Biology Graduate Program, University of Massachusetts Amherst, Amherst, Massachusetts 01003, United States.
J Am Soc Mass Spectrom. 2023 May 3;34(5):931-938. doi: 10.1021/jasms.3c00016. Epub 2023 Apr 4.
The diversity of ubiquitin modifications calls for methods to better characterize ubiquitin chain linkage, length, and morphology. Here, we use multiple linear regression analysis coupled with ion mobility mass spectrometry (IM-MS) to quantify the relative abundance of different ubiquitin dimer isomers. We demonstrate the utility and robustness of this approach by quantifying the relative abundance of different ubiquitin dimers in complex mixtures and comparing the results to the standard, bottom-up ubiquitin AQUA method. Our results provide a foundation for using multiple linear regression analysis and IM-MS to characterize more complex ubiquitin chain architectures.
泛素修饰的多样性要求更好地表征泛素链连接、长度和形态的方法。在这里，我们使用多元线性回归分析结合离子淌度质谱（IM-MS）来定量不同泛素二聚体异构体的相对丰度。我们通过在复杂混合物中定量不同泛素二聚体的相对丰度并将结果与标准的自上而下的泛素 AQUA 方法进行比较，证明了这种方法的实用性和稳健性。我们的结果为使用多元线性回归分析和 IM-MS 来表征更复杂的泛素链结构提供了基础。
