Noncovalent protein tetramers and pentamers with "n" charges yield monomers with n/4 and n/5 charges.
Noncovalent protein tetramers and pentamers with "n" charges yield monomers with n/4 and n/5 charges.
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DOI：
10.1021/ac801883k
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发表时间：
2009-02-15
影响因子：
7.4
通讯作者：
Wysocki, Vicki H.
中科院分区：
文献类型：
--
作者：
Beardsley, Richard L.;Jones, Christopher M.;Galhena, Asiri S.;Wysocki, Vicki H.
近年来，基于质谱的技术已成为表征大分子非共价组装体的结构生物学工具。其中许多工作涉及在质谱仪内保存完整的蛋白质复合物，提供分子量测量，从而确定亚基化学计量和实时监测蛋白质相互作用。人们试图通过将亚基与惰性气体原子（如氩气或氙气）碰撞，将其从完整的复合物中分离出来，从而进一步阐明亚基结构。不幸的是，从这种策略中可以获得的结构信息量受到几乎无处不在的单个展开亚基的喷射的限制。在这里，我们展示了蛋白质-蛋白质复合物碰撞到表面时气相解离的结果。一系列四聚体和五聚体蛋白质的解离表明，通过表面碰撞可以产生通过与气体原子多次碰撞未观察到的替代亚基片段。有证据表明保留了单个亚基结构，并且在某些情况下，保留了亚基和配体之间的非共价相互作用。我们将这些差异归因于离子表面碰撞的快速大能量输入，这导致亚基在单个单体展开之前解离。
In recent years mass spectrometry based techniques have emerged as structural biology tools for the characterization of macromolecular, non-covalent assemblies. Many of these efforts involve preservation of intact protein complexes within the mass spectrometer, providing molecular weight measurements that allow the determination of subunit stoichiometry and real-time monitoring of protein interactions. Attempts have been made to further elucidate subunit architecture through the dissociation of subunits from the intact complex by colliding it into inert gas atoms such as argon or xenon. Unfortunately, the amount of structural information that can be derived from such strategies is limited by the nearly ubiquitous ejection of a single, unfolded subunit. Here, we present results from the gas-phase dissociation of protein-protein complexes upon collision into a surface. Dissociation of a series of tetrameric and pentameric proteins demonstrate that alternative subunit fragments, not observed through multiple collisions with gas atoms, can be generated through surface collision. Evidence is presented for the retention of individual subunit structure, and in some cases, retention of non-covalent interactions between subunits and ligands. We attribute these differences to the rapid large energy input of ion-surface collisions, which leads to the dissociation of subunits prior to the unfolding of individual monomers.
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影响因子：
7.4
作者：
Lancaster, Katherine S.;An, Hyun Joo;Lebrilla, Carlito B.
通讯作者：
Lebrilla, Carlito B.
影响因子：
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作者：
ANTONINI, E
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ANTONINI, E
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作者：
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Superti-Furga, G
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LOO, JA
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LOO, JA
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Badman, ER;Hoaglund-Hyzer, CS;Clemmer, DE
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Clemmer, DE