Following Structural Changes by Thermal Denaturation Using Trapped Ion Mobility Spectrometry-Mass Spectrometry.
Following Structural Changes by Thermal Denaturation Using Trapped Ion Mobility Spectrometry-Mass Spectrometry.
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DOI：
10.1021/acs.jpcb.0c04276
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发表时间：
2020-07-23
期刊：
影响因子：
--
通讯作者：
Fernandez-Lima F
中科院分区：
文献类型：
--
作者：
Jeanne Dit Fouque K;Fernandez-Lima F
作为溶液温度的函数的生物分子的行为通常对于评估其生物活性和功能至关重要。虽然传统上使用光学光谱方法监测生物分子的热诱导变化，但它们的构象变化和解折叠转变仍然具有挑战性。在目前的工作中，牛血清白蛋白（BSA）在天然条件下（100 mM醋酸铵水溶液）的结构转变进行了研究，作为一个函数的起始溶液温度（T = ~23-70 °C）使用的温度控制的纳米电喷雾电离源（nESI）耦合到捕获离子迁移谱-质谱（TIMS-MS）仪器。单体BSA的电荷状态分布从天然样的窄电荷状态（在~23 °C下的[M +12 H]12+ - [M +16 H]16+）和窄迁移率分布变为展开样的宽电荷状态（在~70 °C下高达[M +46 H]46+）和宽迁移率分布。平均电荷状态和碰撞截面（CCS）分布的检查表明存在两种状态的解折叠转变，熔融温度Tm约为56 ± 1 ℃;然而，电荷状态水平下CCS曲线随溶液温度变化的检查显示至少有六种结构转变（T1-T7）。如果起始溶液浓度略微增加（从2 μM增加到25 μM），该方法可以检测早期解离（Td ~34 ± 1 °C）并可能干扰BSA单体熔解曲线的非特异性BSA二聚体和三聚体。在单个实验中，该技术提供了溶液的详细视图，蛋白质结构景观（迁移率vs溶液温度vs每个电荷状态的相对强度）。
The behavior of biomolecules as a function of the solution temperature is often crucial to assess their biological activity and function. While heat-induced changes of biomolecules are traditionally monitored using optical spectroscopy methods, their conformational changes and unfolding transitions remain challenging to interpret. In the present work, the structural transitions of bovine serum albumin (BSA) in native conditions (100 mM aqueous ammonium acetate) were investigated as a function of the starting solution temperature (T = ~23-70 °C) using a temperature-controlled nano-electrospray ionization source (nESI) coupled to a trapped ion mobility spectrometry – mass spectrometry (TIMS-MS) instrument. The charge state distribution of the monomeric BSA changed from a native-like, narrow charge state ([M + 12H]12+ - [M + 16H]16+ at ~23 °C) and narrow mobility distribution towards an unfolded-like, broad charge state (up to [M + 46H]46+ at ~70 °C) and broad mobility distribution. Inspection of the average charge state and collision cross section (CCS) distribution suggested a two-state unfolding transition with a melting temperature Tm ~56 ± 1 °C; however, the inspection of the CCS profiles at the charge state level as a function of the solution temperature showcase at least six structural transitions (T1-T7). If the starting solution concentration is slightly increased (from 2 to 25 μM), this method can detect non-specific BSA dimers and trimers which dissociate early (Td ~34 ± 1 °C) and may disturb the melting curve of the BSA monomer. In a single experiment, this technology provides a detailed view of the solution, protein structural landscape (mobility vs solution temperature vs relative intensity for each charge state).
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影响因子：
2.9
作者：
ANDERSON, DE;BECKTEL, WJ;DAHLQUIST, FW
通讯作者：
DAHLQUIST, FW
影响因子：
7.4
作者：
Benesch, JLP;Sobott, F;Robinson, CV
通讯作者：
Robinson, CV
影响因子：
3.3
作者：
Fouque, Kevin Jeanne Dit;Moreno, Javier;Fernandez-Lima, Francisco
通讯作者：
Fernandez-Lima, Francisco
影响因子：
4.3
作者：
Fouque, Kevin Jeanne Dit;Bisram, Vikash;Fernandez-Lima, Francisco
通讯作者：
Fernandez-Lima, Francisco
影响因子：
1.8
作者：
Butcher, David;Bernad, Sophie;Fernandez-Lima, Francisco
通讯作者：
Fernandez-Lima, Francisco