Electron-capture dissociation and ion mobility mass spectrometry for characterization of the hemoglobin protein assembly
Electron-capture dissociation and ion mobility mass spectrometry for characterization of the hemoglobin protein assembly
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DOI：
10.1002/pro.2712
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发表时间：
2015-08-01
期刊：
影响因子：
8
通讯作者：
Gross, Michael L.
中科院分区：
文献类型：
--
作者：
Cui, Weidong;Zhang, Hao;Gross, Michael L.
天然喷雾具有探测蛋白质组装体的生物物理性质的潜力。在这里，我们报告了一项调查，使用ECD自上而下的测序与FTICR质谱仪和离子迁移率（IM）的测量上的Q-TOF调查碰撞诱导展开的天然样异质四聚体组装，人血红蛋白（hHb），在气相中。据我们所知，这是第一份报告结合ECD和离子迁移率数据在同一个目标蛋白组装描绘碰撞激活对组装大小和碎片的程度和位置的影响。虽然碰撞诱导的血红蛋白组件的展开是清楚地看到IMMS和ECD，后者描绘的区域，越来越多地展开碰撞能量的增加。结果是一致的，与以前的结果均质的蛋白质组装，并加强我们的解释，激活打开的结构的蛋白质组装从灵活的区域，使可用的ECD片段，而不解离的组分蛋白质。
Native spray has the potential to probe biophysical properties of protein assemblies. Here we report an investigation using both ECD top-down sequencing with an FTICR mass spectrometer and ion mobility (IM) measurements on a Q-TOF to investigate the collisionally induced unfolding of a native-like heterogeneous tetrameric assembly, human hemoglobin (hHb), in the gas phase. To our knowledge, this is the first report combining ECD and ion-mobility data on the same target protein assembly to delineate the effects of collisional activation on both assembly size and the extent and location of fragmentation. Although the collision-induced unfolding of the hemoglobin assembly is clearly seen by both IMMS and ECD, the latter delineates the regions that increasingly unfold as the collision energy is increased. The results are consistent with previous outcomes for homogeneous protein assemblies and reinforce our interpretation that activation opens the structure of the protein assembly from the flexible regions to make available ECD fragmentation, without dissociating the component proteins.