193 nm Ultraviolet Photodissociation Mass Spectrometry of Tetrameric Protein Complexes Provides Insight into Quaternary and Secondary Protein Topology.
193 nm Ultraviolet Photodissociation Mass Spectrometry of Tetrameric Protein Complexes Provides Insight into Quaternary and Secondary Protein Topology.
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DOI：
10.1021/jacs.6b03905
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发表时间：
2016-08-31
影响因子：
15
通讯作者：
Brodbelt JS
中科院分区：
文献类型：
--
作者：
Morrison LJ;Brodbelt JS
蛋白质-蛋白质界面和结构对多蛋白质复合物的功能至关重要。基于质谱的技术已经成为表征蛋白质复合物的强有力的策略，特别是对于结构的异质混合物。在本研究中，三个四聚体蛋白质复合物（链霉亲和素，甲状腺素运载蛋白，血红蛋白）在气相中的激活和解离进行了193 nm紫外光解离（UVPD）的表征的高阶结构。高脉冲能量UVPD导致二聚体和低电荷单体的生产，表现出对称的子单元之间的电荷分配（所谓的对称解离途径），与复合物的亚基组织一致。此外，UVPD促进单体亚基的骨架裂解，其丰度对应于蛋白质的更灵活的环区域。
Protein-protein interfaces and architecture are critical to the function of multiprotein complexes. Mass spectrometry-based techniques have emerged as powerful strategies for characterization of protein complexes, particularly for heterogeneous mixtures of structures. In the present study, activation and dissociation of three tetrameric protein complexes (streptavidin, transthyretin, and hemoglobin) in the gas phase was undertaken by 193 nm ultraviolet photodissociation (UVPD) for the characterization of higher order structure. High pulse energy UVPD resulted in the production of dimers and low charged monomers exhibiting symmetrical charge partitioning among the sub-units (the so-called symmetrical dissociation pathways), consistent with the subunit organization of the complexes. In addition, UVPD promoted backbone cleavages of the monomeric subunits, the abundances of which corresponded to the more flexible loop regions of the proteins.