A Dielectric Barrier Discharge Plasma Degrades Proteins to Peptides by Cleaving the Peptide Bond
A Dielectric Barrier Discharge Plasma Degrades Proteins to Peptides by Cleaving the Peptide Bond
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DOI：
10.1007/s11090-019-10053-2
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发表时间：
2019-11-27
影响因子：
3.6
通讯作者：
Bandow, Julia Elisabeth
中科院分区：
文献类型：
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作者：
Krewing, Marco;Schubert, Britta;Bandow, Julia Elisabeth
研究大气压等离子体对蛋白质的影响，以评估等离子体净化的质量，并深入了解等离子体触发的分子事件，这些事件是等离子体医学在细胞、器官和系统水平上进行的观察的基础。据报道，大气压等离子体处理会导致蛋白质降解。然而，降解产物尚未被表征。用 DBD 等离子体处理水溶液中的不同模型蛋白质，我们用不同的方法（布拉德福德测定、应用兰伯特-比尔定律在 280 nm 处的吸收测量、茚三酮测定、尺寸排阻色谱、SDS-PAGE）确认蛋白质发生降解。通过尺寸排阻色谱法检测不同尺寸的肽。茚三酮测定表明肽键被裂解。在羟自由基清除剂 d-甘露醇存在下，等离子体处理最初 10 分钟期间形成的氨基末端浓度降低了 96%，而在较长的处理时间后，甘露醇不再阻止氨基末端的形成，表明羟自由基在蛋白质中肽键的初始裂解中发挥着重要作用，但在裂解所得肽中的肽键中也有其他机制在起作用。肽的产生对血浆净化和血浆医学具有重要意义。验证等离子体净化过程不会产生具有不良特性的蛋白质片段至关重要。在血浆医学中，血浆产生的蛋白质片段可以在治疗的细胞、组织或患者中充当分子触发器，例如以类似蛋白酶的方式调节信号级联。
Effects of atmospheric pressure plasmas on proteins are studied to assess the quality of plasma decontamination and to gain insights into plasma-triggered molecular events underlying observations made in plasma medicine on the cellular, organ, and systemic level. Atmospheric pressure plasma treatment has been reported to cause protein degradation. Degradation products, however, have not been characterized. Treating different model proteins in aqueous solution with a DBD plasma, we confirmed with different methods (Bradford assay, application of Lambert-Beer's law on absorption measurements at 280 nm, ninhydrin assay, size exclusion chromatography, SDS-PAGE) that protein degradation takes place. Peptides of different sizes were detected by size exclusion chromatography. The ninhydrin assay indicated that peptide bonds are cleaved. In the presence of hydroxyl radical scavenger d-mannitol, the concentration of amino termini formed during the initial 10 min of plasma treatment was reduced by 96%, while at longer treatment times mannitol did no longer prevent the formation of amino termini, indicating that hydroxyl radicals play an important role in the initial cleaving of peptide bonds in the protein, but other mechanisms are at play in cleaving the peptide bonds in the resulting peptides. The generation of peptides has implications for plasma decontamination and plasma medicine. It is critical to verify that plasma decontamination processes do not result in protein fragments with undesired properties. In plasma medicine, plasma-generated protein fragments may act as molecular triggers in treated cells, tissues, or patients, e.g., regulating signaling cascades in a protease-like fashion.