PDF(Pubmed)
Abstract：
Native mass spectrometry has recently moved alongside traditional structural biology techniques in its ability to provide clear insights into the composition of protein complexes. However, to date, limited software tools are available for the comprehensive analysis of native mass spectrometry data on protein complexes, particularly for experiments aimed at elucidating the composition of an intact protein complex. Here, we introduce ProSight Native as a start-to-finish informatics platform for analyzing native protein and protein complex data. Combining mass determination via spectral deconvolution with a top-down database search and stoichiometry calculations, ProSight Native can determine the complete composition of protein complexes. To demonstrate its features, we used ProSight Native to successfully determine the composition of the homotetrameric membrane complex Aquaporin Z. We also revisited previously published spectra and were able to decipher the composition of a heterodimer complex bound with two noncovalently associated ligands. In addition to determining complex composition, we developed new tools in the software for validating native mass spectrometry fragment ions and mapping top-down fragmentation data onto three-dimensional protein structures. Taken together, ProSight Native will reduce the informatics burden on the growing field of native mass spectrometry, enabling the technology to further its reach.
摘要：
天然质谱最近与传统的结构生物学技术一起发展，能够提供对蛋白质复合物组成的清晰见解。然而,到目前为止,有限的软件工具可用于蛋白质复合物的天然质谱数据的综合分析，特别是用于旨在阐明完整蛋白质复合物组成的实验。这里,我们引入ProSightNative作为一个开始到完成的信息学平台，用于分析天然蛋白质和蛋白质复合物数据。将通过光谱反卷积进行的质量确定与自上而下的数据库搜索和化学计量计算相结合，ProSightNative可以确定蛋白质复合物的完整组成。为了展示它的特点，我们使用ProSightNative成功地确定了同四聚体膜复合物水通道蛋白Z的组成。我们还重新访问了以前发表的光谱，并且能够破译与两个非共价结合的配体结合的异二聚体复合物的组成。除了确定复杂的成分，我们在软件中开发了新工具，用于验证天然质谱碎片离子，并将自上而下的碎片数据映射到三维蛋白质结构上。一起来看,ProSightNative将减少日益增长的天然质谱领域的信息学负担，使该技术能够进一步发挥作用。
