Fourier-Transform Approach for Reconstructing Macromolecular Mass Defect Profiles
Fourier-Transform Approach for Reconstructing Macromolecular Mass Defect Profiles
复制标题
重建大分子质量缺陷轮廓的傅里叶变换方法
DOI：
10.1021/jasms.1c00317
复制
发表时间：
2022
影响因子：
3.2
通讯作者：
Prell, James S.
中科院分区：
文献类型：
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作者：
Swansiger, Andrew K.;Marty, Michael T.;Prell, James S.
国家的最先进的天然质谱（MS）的方法已被开发用于分析高度异质性的完整的复合物，并提供了许多深入了解的结构和性质的非共价组装，可以很难研究使用变性蛋白质。这些天然MS方法通常可用于研究嵌入洗涤剂胶束、纳米盘和其他膜模拟物中的甚至高度多分散的膜蛋白。然而，表征高度多分散的天然复合物，也是异质性的提出了额外的挑战，原生MS。大分子质量缺陷（MMD）分析的目的是表征异构离子群体混淆加合物的多分散性，并揭示分布的“基地”群众，最近实施的贝叶斯分析软件UniDec。在这里，我们说明了一种替代的，正交MMD分析方法中实现的反卷积程序iFAMS，它利用傅里叶变换（FT）反卷积低分辨率数据与用户输入的参数很少，它可以提供高质量的结果，即使是质谱的信噪比为1.5：1。这种方法，这是基于频域数据，和质量域算法的UniDec之间的协议提供了强有力的证据表明，这两种方法可以准确地表征高度多分散性和异质性的离子种群。FT算法预计将是非常有用的，在表征许多类型的分析物，从膜蛋白到聚合物共轭蛋白质，支链聚合物，和其他大型分析物，以及重建同位素配置文件的高度复杂，但仍然同位素分辨质谱。
State-of-the-art native mass spectrometry (MS) methods have been developed for analysis of highly heterogeneous intact complexes and have provided much insight into the structure and properties of noncovalent assemblies that can be difficult to study using denatured proteins. These native MS methods can often be used to study even highly polydisperse membrane proteins embedded in detergent micelles, nanodiscs, and other membrane mimics. However, characterizing highly polydisperse native complexes which are also heterogeneous presents additional challenges for native MS. Macromolecular mass defect (MMD) analysis aims to characterize heterogeneous ion populations obfuscated by adduct polydispersity and reveal the distribution of “base” masses, and was recently implemented in the Bayesian analysis software UniDec. Here, we illustrate an alternative, orthogonal MMD analysis method implemented in the deconvolution program iFAMS, which takes advantage of Fourier transform (FT) to deconvolve low-resolution data with few user-input parameters and which can provide high quality results even for mass spectra with a signal-to-noise ratio of ∼5:1. Agreement between this method, which is based on frequency-domain data, and the mass-domain algorithm of UniDec provides strong evidence that both methods can accurately characterize highly polydisperse and heterogeneous ion populations. The FT algorithm is expected to be very useful in characterizing many types of analytes ranging from membrane proteins to polymer-conjugated proteins, branched polymers, and other large analytes, as well as for reconstructing isotope profiles for highly complex but still isotope-resolved mass spectra.