Discovery of the Xenon-Protein Interactome Using Large-Scale Measurements of Protein Folding and Stability.
Discovery of the Xenon-Protein Interactome Using Large-Scale Measurements of Protein Folding and Stability.
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DOI：
10.1021/jacs.1c11900
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发表时间：
2022-03-09
影响因子：
15
通讯作者：
Fitzgerald, Michael C.
中科院分区：
文献类型：
--
作者：
Wiebelhaus, Nancy;Singh, Niven;Zhang, Peng;Craig, Stephen L.;Beratan, David N.;Fitzgerald, Michael C.
惰性气体在生物系统中的分子间相互作用与许多生化反应有关，包括细胞凋亡、炎症、麻醉、镇痛和神经保护。这些反应背后的分子作用模式在很大程度上是未知的。这在很大程度上是由于研究蛋白质-气体相互作用的实验技术有限。少数几种适用于此类研究的技术是相对较低的吞吐量，并且需要大量纯化的蛋白质。因此，它们不能进行对蛋白质靶标发现有用的大规模分析。在这里，我们报告了氧化速率蛋白质稳定性(SPROX)和有限蛋白质分解(LIP)方法的应用，通过蛋白质折叠稳定性测量来检测蛋白质组尺度上的蛋白质-Xe相互作用。用SPROX和LIP技术分析了酵母蛋白质组中5,000多个蛋氨酸多肽和5,000多个半胰蛋白酶多肽，分别定位于~1,500和~950个蛋白质。SPROX和LIP分析分别鉴定出31种和60种Xe相互作用蛋白，其中没有一种以前已知与Xe结合。对蛋白质组学结果的生物信息学分析表明，这些Xe相互作用的蛋白质在参与ATP驱动的过程中富含。已确定的一小部分蛋白质靶标与先前建立的与氙气的麻醉和器官保护特性有关的作用模式有关。这些结果丰富了我们对与生物相关的氙气相互作用的认识和理解。这里开发的氙气样品制备方案和分析方法也普遍适用于在复杂的生物混合物中发现广泛的其他蛋白质-气体相互作用，如细胞裂解物。
The intermolecular interactions of noble gases in biological systems are associated with numerous biochemical responses, including apoptosis, inflammation, anesthesia, analgesia, and neuroprotection. The molecular modes of action underlying these responses are largely unknown. This is in large part due to the limited experimental techniques to study protein-gas interactions. The few techniques that are amenable to such studies are relatively low throughput and require large amounts of purified proteins. Thus, they do not enable the large-scale analyses that are useful for protein-target discovery. Here we report the application of Stability of Proteins from Rates of Oxidation (SPROX) and limited proteolysis (LiP) methodologies to detect protein-xenon interactions on the proteomic scale using protein folding stability measurements. Over 5,000 methionine-containing peptides and over 5,000 semi-tryptic peptides, mapping to ~1,500 and ~950 proteins, respectively, in the yeast proteome, were assayed for Xe-interacting activity using the SPROX and LiP techniques. The SPROX and LiP analyses identified 31 and 60 Xe-interacting proteins, respectively, none of which were previously known to bind Xe. A bioinformatics analysis of the proteomic results revealed that these Xe-interacting proteins were enriched in those involved in ATP-driven processes. A fraction of the protein targets that were identified is tied to previously established modes of action related to xenon’s anesthetic and organoprotective properties. These results enrich our knowledge and understanding of biologically relevant xenon interactions. The sample preparation protocols and analytical methodologies developed here for xenon are also generally applicable to the discovery of a wide range of other protein-gas interactions in complex biological mixtures, such as cell lysates.
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影响因子：
14.9
作者：
Gene Ontology Consortium
通讯作者：
Gene Ontology Consortium
影响因子：
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作者：
DeArmond, Patrick D.;Xu, Ying;Strickland, Erin C.;Daniels, Kyle G.;Fitzgerald, Michael C.
通讯作者：
Fitzgerald, Michael C.
影响因子：
15
作者：
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通讯作者：
Luthey-Schulten, Zaida
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作者：
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通讯作者：
Dickinson, Robert
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作者：
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