Effect of DMSO on Protein Structure and Interactions Assessed by Collision-Induced Dissociation and Unfolding
Effect of DMSO on Protein Structure and Interactions Assessed by Collision-Induced Dissociation and Unfolding
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DOI：
10.1021/acs.analchem.7b02329
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发表时间：
2017-09-19
影响因子：
7.4
通讯作者：
Abell, Chris
中科院分区：
文献类型：
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作者：
Chan, Daniel S. -H.;Kavanagh, Madeline E.;Abell, Chris
鉴于DMSO在生物化学和生物物理测定中的频繁使用，需要了解DMSO浓度对蛋白质解离或解折叠行为的影响。在这项研究中，DMSO对亲和素和结核分枝杆菌（Mtb）CYP 142 A1的结构和相互作用的影响进行了评估，通过碰撞诱导解离（CID）和碰撞诱导解折叠（CIU），通过纳米电喷雾离子迁移率质谱（nESI-IM-MS）监测。DMSO浓度高于4%（v/v）通过其对电荷状态分布（CSD）以及在单个电荷状态水平的影响使抗生物素蛋白四聚体朝向解离和展开不稳定。相比之下，DMSO既防止血红素损失，又增加了CYP 142 A1的稳定性，即使在高达40%的DMSO下也能解折叠。串联MS/MS实验表明，DMSO可以改变CYP 142 A1的解离途径，而CIU揭示了血红素基团对CYP 142 A1结构的保护作用。
Given the frequent use of DMSO in biochemical and biophysical assays, it is desirable to understand the influence of DMSO concentration on the dissociation or unfolding behavior of proteins. In this study, the effects of DMSO on the structure and interactions of avidin and Mycobacterium tuberculosis (Mtb) CYP142A1 were assessed through collision induced dissociation (CID) and collision-induced unfolding (CIU) as monitored by nanoelectrospray ionization-ion mobility-mass spectrometry (nESI-IM-MS). DMSO concentrations higher than 4% (v/v) destabilize the avidin tetramer toward dissociation and unfolding, via both its effects on charge state distribution (CSD) as well as at the level of individual charge states. In contrast, DMSO both protects against heme loss and increases the stability of CYP142A1 toward unfolding even up to 40% DMSO. Tandem MS/MS experiments showed that DMSO could modify the dissociation pathway of CYP142A1, while CIU revealed the protective effect of the heme group on the structure of CYP142A1.