Conformational Landscapes of Ubiquitin, Cytochrome c, and Myoglobin: Uniform Field Ion Mobility Measurements in Helium and Nitrogen Drift Gas.
Conformational Landscapes of Ubiquitin, Cytochrome c, and Myoglobin: Uniform Field Ion Mobility Measurements in Helium and Nitrogen Drift Gas.
复制标题
DOI：
10.1016/j.ijms.2017.09.014
复制
发表时间：
2018-04
影响因子：
1.8
通讯作者：
McLean JA
中科院分区：
文献类型：
--
作者：
May JC;Jurneczko E;Stow SM;Kratochvil I;Kalkhof S;McLean JA
在这项研究中，利用商业均匀场漂移管离子迁移率质谱仪（IM-MS）测量三种充分研究的蛋白质的气相构象群体：泛素（8566 Da），细胞色素c（12，359 Da），以及脱辅基蛋白和全蛋白形式的肌红蛋白（分别为16，951和17，567 Da），以评估该技术在大规模结构蛋白质组学应用中的用途。从酸性有机（pH ~3）或水性缓冲（pH ~6.6）溶液相条件电喷雾蛋白质，其产生对应于扩展（未折叠）和紧凑（折叠）气相构象群体的宽范围的阳离子电荷状态。相应的碰撞截面（CCS）的测量进行了编译显着的离子迁移率峰在每个电荷状态下观察到的功能，以映射这些蛋白质在氦气和氮气漂移气体的构象景观。据观察，构象景观是相似的，在两种漂移气体，与差异主要归因于离子加热氦操作过程中，由于操作的仪器具有较高的压差的必要性。在氮中观察到更高的分辨能力，这使得紧密间隔的构象异构体群体的结构分辨率略好。仪器被发现是特别善于测量低丰度构象，这是唯一存在于温和的条件下，最大限度地减少离子加热。这项工作代表了迄今为止这三种蛋白质的最大的单一离子迁移率CCS调查，具有266个CCS值和117个离子迁移率谱，其中许多以前没有报道过。
In this study, a commercial uniform field drift tube ion mobility-mass spectrometer (IM-MS) was utilized to measure the gas-phase conformational populations of three well-studied proteins: ubiquitin (8566 Da), cytochrome c (12,359 Da), and myoglobin in both apo and holo forms (16,951 and 17,567 Da, respectively) in order to evaluate the use of this technology for broadscale structural proteomics applications. Proteins were electrosprayed from either acidic organic (pH ~3) or aqueous buffered (pH ~6.6) solution phase conditions, which generated a wide range of cation charge states corresponding to both extended (unfolded) and compact (folded) gas-phase conformational populations. Corresponding collision cross section (CCS) measurements were compiled for significant ion mobility peak features observed at each charge state in order to map the conformational landscapes of these proteins in both helium and nitrogen drift gases. It was observed that the conformational landscapes were similar in both drift gases, with differences being attributed primarily to ion heating during helium operation due to the necessity of operating the instrument with higher pressure differentials. Higher resolving powers were observed in nitrogen, which allowed for slightly better structural resolution of closely-spaced conformer populations. The instrumentation was found to be particularly adept at measuring low abundance conformers which are only present under gentle conditions which minimize ion heating. This work represents the single largest ion mobility CCS survey published to date for these three proteins with 266 CCS values and 117 ion mobility spectra, many of which have not been previously reported.
登录
查看更多内容
影响因子：
1.8
作者：
Bleiholder, Christian;Contreras, Stephanie;Bowers, Michael T.
通讯作者：
Bowers, Michael T.
影响因子：
1.8
作者：
Bleiholder, Christian;Wyttenbach, Thomas;Bowers, Michael T.
通讯作者：
Bowers, Michael T.
DOI：
10.1016/j.jasms.2006.06.005
发表时间：
2006-09-01
影响因子：
3.2
作者：
Ibrahim, Yehia;Tang, Keqi;Smith, Richard D.
通讯作者：
Smith, Richard D.
DOI：
10.1016/1044-0305(93)85025-s
发表时间：
1993-08-01
影响因子：
3.2
作者：
COVEY, T;DOUGLAS, DJ
通讯作者：
DOUGLAS, DJ
影响因子：
7.4
作者：
Clowers, Brian H.;Belov, Mikhail E.;Smith, Richard D.
通讯作者：
Smith, Richard D.