Plasma-Generated OH Radical Production for Analyzing Three-Dimensional Structure in Protein Therapeutics
Plasma-Generated OH Radical Production for Analyzing Three-Dimensional Structure in Protein Therapeutics
复制标题
等离子体产生的 OH 自由基产生用于分析蛋白质治疗中的三维结构
DOI：
10.1038/s41598-017
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
Minkoff BB, Blatz JM
中科院分区：
文献类型：
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作者：
Minkoff BB, Blatz JM
蛋白质三维结构在溶液中动态变化取决于配体和相互作用蛋白质的存在。检测这些蛋白质构象变化的方法包括使用质谱法的“蛋白质足迹”。我们在此描述了一种新技术，PLIMB（等离子体诱导生物分子修饰），它可以从水中产生µs的羟基自由基爆发，通过改变氨基酸侧链的溶剂可及性来测量蛋白质结构的变化。PLIMB首先以模型化合物为基准，然后应用于一个生物学问题，即配体（EGF）诱导表皮生长因子受体（EGFR）外（外）结构域构象的变化。加入EGF后氧化减少的区域沿着二聚化界面下降，与晶体结构的模型一致。这些结果表明，血浆中从水中产生的羟基自由基可以用来绘制蛋白质的构象变化，并提供了一种容易获得的方法来研究溶液中的蛋白质结构。
Protein three-dimensional structure dynamically changes in solution depending on the presence of ligands and interacting proteins. Methods for detecting these changes in protein conformation include ‘protein footprinting,’ using mass spectrometry. We describe herein a new technique, PLIMB (Plasma Induced Modification of Biomolecules), that generates µs bursts of hydroxyl radicals from water, to measure changes in protein structure via altered solvent accessibility of amino acid side chains. PLIMB was first benchmarked with model compounds, and then applied to a biological problem, i.e., ligand (EGF) induced changes in the conformation of the external (ecto) domain of Epidermal Growth Factor Receptor (EGFR). Regions in which oxidation decreased upon adding EGF fall along the dimerization interface, consistent with models derived from crystal structures. These results demonstrate that plasma-generated hydroxyl radicals from water can be used to map protein conformational changes, and provide a readily accessible means of studying protein structure in solution.