Revealing the Quaternary Structure of a Heterogeneous Noncovalent Protein Complex through Surface-Induced Dissociation
Revealing the Quaternary Structure of a Heterogeneous Noncovalent Protein Complex through Surface-Induced Dissociation
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DOI：
10.1021/ac200452b
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发表时间：
2011-04-15
影响因子：
7.4
通讯作者：
Wysocki, Vicki H.
中科院分区：
文献类型：
--
作者：
Blackwell, Anne E.;Dodds, Eric D.;Wysocki, Vicki H.
随着科学家开始了解四级结构促进蛋白质功能的程度，确定非共价蛋白质复合物内的亚基排列变得越来越重要。虽然原生质谱法显示出对非共价复合物的研究的希望，但对亚基结构的确定几乎没有进展，并且没有质谱活化方法产生完整的拓扑结构信息。在这里，我们说明了表面诱导的异源六聚体，丰卡霉素腈水合酶，直接分解成其组成的三聚体。我们提出，这种激活与高能量沉积相结合的单步性质允许在显著的展开或其他大规模重排之前解离。这种方法可以潜在地允许蛋白质复合物解离成亚复合物，促进亚基接触的映射，从而确定蛋白质复合物的四级结构。
As scientists begin to appreciate the extent to which quaternary structure facilitates protein function, determination of the subunit arrangement within noncovalent protein complexes is increasingly important. While native mass spectrometry shows promise for the study of noncovalent complexes, few developments have been made toward the determination of subunit architecture, and no mass spectrometry activation method yields complete topology information. Here, we illustrate the surface-induced dissociation of a heterohexamer, toyocamycin nitrile hydratase, directly into its constituent trimers. We propose that the single-step nature of this activation in combination with high energy deposition allows for dissociation prior to significant unfolding or other large-scale rearrangement This method can potentially allow for dissociation of a protein complex into subcomplexes, facilitating the mapping of subunit contacts and thus determination of quaternary structure of protein complexes.