Enhanced Characterization of Membrane Protein Complexes by Ultraviolet Photodissociation Mass Spectrometry
Enhanced Characterization of Membrane Protein Complexes by Ultraviolet Photodissociation Mass Spectrometry
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DOI：
10.1021/acs.analchem.9b03689
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发表时间：
2020-01-07
影响因子：
7.4
通讯作者：
Brodbelt, Jennifer S.
中科院分区：
文献类型：
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作者：
Sipe, Sarah N.;Patrick, John W.;Brodbelt, Jennifer S.
开发化学伴侣以溶解水溶液中的膜蛋白复合物，可以对其类天然组件进行气相分析，包括快速评估稳定性和相互作用的伙伴。然而，迄今为止，蛋白质一级序列的表征仍然有限。紫外线光解 (UVPD) 为大肠杆菌氨通道 (AmtB) 生成大量序列离子，改进了水通道蛋白 Z (AqpZ) 可能的翻译后修饰的定位，并超越了之前关于大电导机械敏感通道 (MscL) 序列覆盖的报道。 UVPD 序列离子丰度的变化已被证明与某些扰动引起的结构变化相对应。本文报道了初步结果，以阐明当与各种磷脂结合时，MscL 的刚性或灵活性增加。
Development of chemical chaperones to solubilize membrane protein complexes in aqueous solutions has allowed for gas-phase analysis of their native-like assemblies, including rapid evaluation of stability and interacting partners. Characterization of protein primary sequence, however, has thus far been limited. Ultraviolet photodissociation (UVPD) generates a multitude of sequence ions for the E. coli ammonia channel (AmtB), provides improved localization of a possible post-translational modification of aquaporin Z (AqpZ), and surpasses previous reports of sequence coverage for mechanosensitive channel of large conductance (MscL). Variations in UVPD sequence ion abundance have been shown to correspond to structural changes induced upon some perturbation. Preliminary results are reported here for elucidating increased rigidity or flexibility of MscL when bound to various phospholipids.