Characterization of Intramolecular Interactions of Cytochrome c Using Hydrogen-Deuterium Exchange-Trapped Ion Mobility Spectrometry-Mass Spectrometry and Molecular Dynamics.
Characterization of Intramolecular Interactions of Cytochrome c Using Hydrogen-Deuterium Exchange-Trapped Ion Mobility Spectrometry-Mass Spectrometry and Molecular Dynamics.
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DOI：
10.1021/acs.analchem.7b00844
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发表时间：
2017-09-05
影响因子：
7.4
通讯作者：
Fernandez-Lima F
中科院分区：
文献类型：
--
作者：
Molano-Arevalo JC;Jeanne Dit Fouque K;Pham K;Miksovska J;Ridgeway ME;Park MA;Fernandez-Lima F
球状蛋白质，如细胞色素c（cyt c），显示出有组织的天然构象，由氢键相互作用网络维持。在本工作中，通过关联离子中性碰撞截面（CCS）和电荷状态与起始溶液条件和脱溶剂后的时间，使用碰撞诱导活化（CIA），时间分辨氢/氘反交换（HDX）和捕获离子迁移谱-质谱（TIMS-MS）的动力学捕获的中间体的结构查询。TIMS分析仪的高离子迁移率分辨能力允许识别新的离子迁移率带，在+6至+21电荷状态下产生总共63个迁移率带，在-5至-10电荷状态下产生20个迁移率带。迁移率选择HDX率表明，对于相同的电荷状态，具有较大CCS的构象呈现更快的HDX率在正离子和负离子模式，这表明电荷网站和邻近的交换网站上的可及表面积定义的交换率，而不管电荷状态。互补分子动力学模拟允许产生候选结构和从天然（N）到熔融球（MG）到动力学中间体（U）途径的折叠过渡的机械模型。我们的研究结果表明，细胞色素C的主要结构展开与N-和C-末端螺旋的距离和随后的溶剂暴露的疏水性，含血红素腔。
Globular proteins, such as cytochrome c (cyt c), display an organized native conformation, maintained by a hydrogen bond interaction network. In the present work, the structural interrogation of kinetically trapped intermediates of cyt c was performed by correlating the ion-neutral collision cross section (CCS) and charge state with the starting solution conditions and time after desolvation using collision induced activation (CIA), time resolved hydrogen/deuterium back exchange (HDX) and trapped ion mobility spectrometry - mass spectrometry (TIMS-MS). The high ion mobility resolving power of the TIMS analyzer allowed the identification of new ion mobility bands, yielding a total of 63 mobility bands over the +6 to +21 charge states and 20 mobility bands over the −5 to −10 charge states. Mobility selected HDX rates showed that for the same charge state, conformers with larger CCS present faster HDX rates in both positive and negative ion mode, suggesting that the charge sites and neighboring exchange sites on the accessible surface area define the exchange rate regardless of the charge state. Complementary molecular dynamic simulations permitted the generation of candidate structures and a mechanistic model of the folding transitions from native (N) to molten globule (MG) to kinetic intermediates (U) pathways. Our results suggest that cyt c major structural unfolding is associated with the distancing of the N- and C- terminal helices and subsequent solvent exposure of the hydrophobic, heme-containing cavity.
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DOI：
10.1007/s12127-016-0198-z
发表时间：
2016-09-01
影响因子：
--
作者：
Adams, Kendra J.;Montero, Dennise;Fernandez-Lima, Francisco
通讯作者：
Fernandez-Lima, Francisco
影响因子：
7.4
作者：
Badman, ER;Hoaglund-Hyzer, CS;Clemmer, DE
通讯作者：
Clemmer, DE
DOI：
10.1039/c6cp04418b
发表时间：
2016-09-29
期刊：
Physical chemistry chemical physics : PCCP
影响因子：
--
作者：
Garabedian A;Butcher D;Lippens JL;Miksovska J;Chapagain PP;Fabris D;Ridgeway ME;Park MA;Fernandez-Lima F
通讯作者：
Fernandez-Lima F
影响因子：
1.6
作者：
Fernandez-Lima, F. A.;Kaplan, D. A.;Park, M. A.
通讯作者：
Park, M. A.
影响因子：
2.9
作者：
Fernandez-Lima, F. A.;Wei, H.;Russell, D. H.
通讯作者：
Russell, D. H.