Symmetry of 4-Oxalocrotonate Tautomerase Trimers Influences Unfolding and Fragmentation in the Gas Phase.
Symmetry of 4-Oxalocrotonate Tautomerase Trimers Influences Unfolding and Fragmentation in the Gas Phase.
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DOI：
10.1021/jacs.2c03564
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发表时间：
2022-07-13
影响因子：
15
通讯作者：
Brodbelt, Jennifer S.
中科院分区：
文献类型：
--
作者：
Sipe, Sarah N.;Lancaster, Emily B.;Butalewicz, Jamie P.;Whitman, Christian P.;Brodbelt, Jennifer S.
最近在互变酶超家族（TSF）中发现的三聚体的不对称排列为这个已经具有机械多样性的超家族增加了结构多样性。不对称三聚体的分类以前已经用x射线晶体学确定。在这里，采用天然质谱（MS）和紫外光解（UVPD）作为一种综合策略，以更快速和敏感地区分对称和不对称三聚体。具体而言，利用UVPD研究了碰撞加热引发的对称和不对称三聚体的展开，揭示了独特的气相展开途径。UVPD模式从天然的紧凑三聚体结构到展开的扩展构象的变化表明，不对称三聚体中的高阶结构重排被认为是由盐桥三聚体稳定的，而对称三聚体中没有盐桥三聚体。因此，发现对称三聚体在气相中稳定性较差，导致UVPD碎片总体上增强，并且基于蛋白质片段氢迁移程度的高阶重构显着差异。不对称三聚体稳定性的提高可能证明了它们的进化和随之而来的TSF多样化是合理的。将TSF成员分类为对称或非对称三聚体有助于描述TSF的进化史。
The recent discovery of asymmetric arrangements of trimers in the tautomerase superfamily (TSF) adds structural diversity to this already mechanistically diverse superfamily. Classification of asymmetric trimers has previously been determined using X-ray crystallography. Here, native mass spectrometry (MS) and ultraviolet photodissociation (UVPD) are employed as an integrated strategy for more rapid and sensitive differentiation of symmetric and asymmetric trimers. Specifically, the unfolding of symmetric and asymmetric trimers initiated by collisional heating was probed using UVPD, which revealed unique gas-phase unfolding pathways. Variations in UVPD patterns from native-like, compact trimeric structures to unfolded, extended conformations indicate a rearrangement of higher-order structure in the asymmetric trimers that are believed to be stabilized by salt-bridge triads, which are absent from the symmetric trimers. Consequently, the symmetric trimers were found to be less stable in the gas phase, resulting in enhanced UVPD fragmentation overall and a notable difference in higher-order re-structuring based on the extent of hydrogen migration of protein fragments. The increased stability of the asymmetric trimers may justify their evolution and concomitant diversification of the TSF. Facilitating the classification of TSF members as symmetric or asymmetric trimers assists in delineating the evolutionary history of the TSF.
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DOI：
10.1074/jbc.m117.815340
发表时间：
2018-02-16
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Davidson R;Baas BJ;Akiva E;Holliday GL;Polacco BJ;LeVieux JA;Pullara CR;Zhang YJ;Whitman CP;Babbitt PC
通讯作者：
Babbitt PC
影响因子：
7.4
作者：
McCabe JW;Shirzadeh M;Walker TE;Lin CW;Jones BJ;Wysocki VH;Barondeau DP;Clemmer DE;Laganowsky A;Russell DH
通讯作者：
Russell DH
一种集成的天然质谱和自上而下的蛋白质组学方法，该方法将序列连接到大分子复合物的结构和功能。
影响因子：
21.8
作者：
Li H;Nguyen HH;Ogorzalek Loo RR;Campuzano IDG;Loo JA
通讯作者：
Loo JA
影响因子：
12.4
作者：
通讯作者：
--
影响因子：
2.9
作者：
Baas, Bert-Jan;Medellin, Brenda P.;Whitman, Christian P.
通讯作者：
Whitman, Christian P.