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A mass spectrometry-based approach gives new insight into organotin-protein interactions.
Metallomics ( IF 3.4 ) Pub Date : 2020-09-09 , DOI: 10.1039/d0mt00171f
Jonas M Will 1 , Catharina Erbacher , Michael Sperling , Uwe Karst
Affiliation  

In this study, the combination of speciation analysis and native mass spectrometry is presented as a powerful tool to gain new insight into the diverse interactions of environmentally relevant organotin compounds (OTCs) with proteins. Analytical standards of model proteins, such as β-lactoglobulin A (LGA), were thereby incubated with different phenyl- and butyltins. For adduct identification and characterization, the incubated samples were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) and electrospray ionization-mass spectrometry (ESI-MS) in combination with size exclusion chromatography (SEC). It allowed for a mild separation, which was most crucial to preserve the acid-labile organotin–protein adducts during their analyses. The binding of triorganotin compounds, such as triphenyltin, was shown to be sulfhydryl-directed by using cysteine-specific protein labeling. However, the sole availability of reduced cysteine residues in proteins did not automatically enable adduct formation. This observation complements previous studies and indicates the necessity of a highly specific binding pocket, which was identified for the model protein LGA via enzymatic digestion experiments. In contrast to triorganotins, their natural di- and mono-substituted degradation products, such as dibutyltin, revealed to be less specific regarding their binding to several proteins. Further, it also did not depend on reduced cysteine residues within the protein. In this context, they can probably act as linker molecules, interconnecting proteins, and leading to dimers and probably to higher oligomers. Furthermore, dibutyltin was observed to induce hydrolysis of the protein's peptide backbone at a specific site. Concerning unknown long-term toxic effects, our studies emphasize the importance of future studies on di- and mono-substituted OTCs.

中文翻译:

基于质谱的方法为有机锡-蛋白质相互作用提供了新的见解。

在这项研究中,形态分析和天然质谱的结合是一种强大的工具,可以对环境相关的有机锡化合物 (OTC) 与蛋白质的各种相互作用进行新的了解。模型蛋白质的分析标准,例如 β-乳球蛋白 A (LGA),因此与不同的苯基锡和丁基锡一起孵育。对于加合物的鉴定和表征,通过电感耦合等离子体质谱 (ICP-MS) 和电喷雾电离质谱 (ESI-MS) 结合尺寸排阻色谱 (SEC) 对孵育的样品进行分析。它允许温和的分离,这对于在分析过程中保留酸不稳定的有机锡-蛋白质加合物至关重要。三有机锡化合物的结合,如三苯基锡,通过使用半胱氨酸特异性蛋白质标记显示出是巯基导向的。然而,蛋白质中减少的半胱氨酸残基的唯一可用性并不能自动形成加合物。这一观察结果补充了之前的研究,并表明了高度特异性结合口袋的必要性,该口袋是为模型蛋白 LGA 确定的通过酶消化实验。与三有机锡相比,它们的天然二取代和单取代降解产物,如二丁基锡,表明它们与几种蛋白质的结合特异性较低。此外,它也不依赖于蛋白质内减少的半胱氨酸残基。在这种情况下,它们可能充当连接分子,互连蛋白质,并导致二聚体和更高的寡聚体。此外,观察到二丁基锡在特定位点诱导蛋白质肽骨架的水解。关于未知的长期毒性作用,我们的研究强调了未来对双取代和单取代 OTC 研究的重要性。
更新日期:2020-11-03
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