Dissociation of multiple protein ion charge states following a single gas-phase purification and concentration procedure
Dissociation of multiple protein ion charge states following a single gas-phase purification and concentration procedure
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DOI：
10.1021/ac025587
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发表时间：
2002-09-15
影响因子：
7.4
通讯作者：
McLuckey, SA
中科院分区：
文献类型：
--
作者：
He, M;Reid, GE;McLuckey, SA
从一个单一的浓缩和纯化的电荷状态，然后通过激活每个电荷状态的前体离子电荷状态的范围的形成，被引入作为一种手段，以获得更多的蛋白质结构信息比从一个单一的电荷状态单独解离。该方法使用在[M +8H]（8+）电荷状态的浓缩和纯化之后的噬菌体MS 2病毒外壳蛋白的[M +8H]（8+）到[M +6 H]（6+）前体离子的非共振碰撞活化来说明。在离子阱碰撞活化研究[M +12 H]（12+）解离为[M +5 H]（5+）离子的过程中，根据前体离子电荷状态的影响选择了电荷状态的范围。气相离子/离子质子转移反应和离子停车技术被施加到纯化和浓缩选定的前体离子的电荷状态，以及简化的产品离子光谱。高电荷态离子的片段优先在N-末端侧的脯氨酸残基，而最低电荷态的研究的产物离子光谱主要由C-末端天冬氨酸裂解。最大的结构信息是通过分裂的中间电荷状态。
The formation of a range of precursor ion charge states from a single concentrated and purified charge state, followed by activation of each charge state, is introduced as a means to obtain more protein structural information than is available from dissociation of a single charge state alone. This approach is illustrated using off-resonance collisional activation of the [M + 8H](8+) to [M + 6H](6+) precursor ions of the bacteriophage MS2 viral coat protein following concentration and purification of the [M + 8H](8+) charge state. Ibis range of charge states was selected on the basis of an ion trap collisional activation study of the effects of precursor ion charge state on the dissociation of the [M + 12H](12+) to [M + 5H](5+) ions. Gas-phase ion/ ion proton-transfer reactions and the ion parking technique were applied to purify and concentrate selected precursor ion charge states as well as to simplify the product ion spectra. The high-charge-state ions fragment preferentially at the N-terminal side of proline residues while the product ion spectra of the lowest charge states investigated are dominated by C-terminal aspartic acid cleavages. Maximum structural information is obtained by fragmentation of the intermediate-charge states.