Native mass spectrometry imaging of intact proteins and protein complexes in thin tissue sections
Native mass spectrometry imaging of intact proteins and protein complexes in thin tissue sections
复制标题
DOI：
10.1016/j.ijms.2017.10.009
复制
发表时间：
2019-03-01
影响因子：
1.8
通讯作者：
Cooper, Helen J.
中科院分区：
文献类型：
--
作者：
Griffiths, Rian L.;Sisley, Emma K.;Cooper, Helen J.
在这里，我们提出了原生液体萃取表面分析（LESA）质谱成像的蛋白质和蛋白质复合物从小鼠大脑和肝脏组织。在特征性低电荷状态下检测到完整的蛋白质，表明蛋白质保持折叠。在大脑中，丰富的蛋白质，如泛素和β胸腺素4在整个组织中被均匀地检测到，而其他蛋白质，如神经颗粒蛋白，则定位于特定的解剖区域。在肝脏中，我们展示了蛋白质复合物（四聚血红蛋白）以及脂肪酸结合蛋白的成像。有趣的是，天然样溶剂的使用使得能够提取蛋白质，这在先前使用变性溶剂的LESA实验中没有观察到，即，天然LESA可用于扩大观察到的蛋白质的范围。我们还提出了本地LESA离子迁移谱，并表明，从组织中提取的蛋白质的碰撞截面可以通过行波离子迁移谱来确定。泛素的5+离子的碰撞截面计算为1047埃（2），与泛素蛋白质标准溶液的测量结果非常一致。还计算了β-胸腺素4、β-胸腺素10和两种未鉴定蛋白质的4+离子的碰撞截面，以及分子量为15660 Da的未鉴定蛋白质的10+离子的碰撞截面。（C）2017作者。由爱思唯尔公司出版
Here, we present native liquid extraction surface analysis (LESA) mass spectrometry imaging of proteins and protein complexes from mouse brain and liver tissue. Intact proteins were detected in characteristically low charge states, indicating that the proteins remain folded. In brain, abundant proteins such as ubiquitin and beta thymosin 4 were detected homogeneously across the tissue whereas other proteins, such as neurogranin, were localised in specific anatomical regions. In liver, we demonstrate imaging of a protein complex (tetrameric hemoglobin), as well as fatty acid binding protein. Interestingly, the use of native-like solvents enables extraction of proteins which have not previously been observed in LESA experiments employing denaturing solvents, i.e., native LESA can be applied to extend the range of proteins observed. We also present native LESA ion mobility spectrometry and show that the collision cross sections of proteins extracted from tissue may be determined by travelling wave ion mobility spectrometry. The collision cross section of the 5+ ion of ubiquitin was calculated as 1047 angstrom(2), in good agreement with measurements of ubiquitin protein standard solutions. Collision cross sections for the 4+ ions of beta-thymosin 4, beta-thymosin 10 and two unidentified proteins were also calculated, together with that of a 10+ ion of an unidentified protein of molecular weight 15660 Da. (C) 2017 The Authors. Published by Elsevier B.V.