Native Ambient Mass Spectrometry of an Intact Membrane Protein Assembly and Soluble Protein Assemblies Directly from Lens Tissue
Native Ambient Mass Spectrometry of an Intact Membrane Protein Assembly and Soluble Protein Assemblies Directly from Lens Tissue
复制标题
对直接来自晶状体组织的完整膜蛋白组装体和可溶性蛋白组装体进行天然环境质谱分析
DOI：
10.1002/ange.202201458
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发表时间：
2022
影响因子：
--
通讯作者：
Hale O
中科院分区：
文献类型：
--
作者：
Hale O
膜蛋白约占治疗靶点的三分之二，但由于其丰度和溶解度低，对结构分析提出了重大挑战。现有的结构分析方法依赖于膜蛋白的过表达和/或纯化，从而消除了与实际生理环境的任何联系。在这里，我们展示了直接从组织中获得的完整寡聚膜蛋白的质谱分析。水通道蛋白-0以113 kDa四聚体形式存在，每个亚基具有六个跨膜螺旋。 我们报告了直接从羊眼透镜切片（未进行样品预处理）中获得的完整组装件的表征。通过四聚体和亚基的质量测量以及自顶向下质谱法确认蛋白质身份，并通过质谱成像确定空间分布。我们的方法允许同时分析组织中的可溶性蛋白质组装。
Membrane proteins constitute around two‐thirds of therapeutic targets but present a significant challenge for structural analysis due to their low abundance and solubility. Existing methods for structural analysis rely on over‐expression and/or purification of the membrane protein, thus removing any links back to actual physiological environment. Here, we demonstrate mass spectrometry analysis of an intact oligomeric membrane protein directly from tissue. Aquaporin‐0 exists as a 113 kDa tetramer, with each subunit featuring six transmembrane helices. We report the characterisation of the intact assembly directly from a section of sheep eye lens without sample pre‐treatment. Protein identity was confirmed by mass measurement of the tetramer and subunits, together with top‐down mass spectrometry, and the spatial distribution was determined by mass spectrometry imaging. Our approach allows simultaneous analysis of soluble protein assemblies in the tissue.