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首页> 外文期刊>Chemistry: A European journal >A nanoporous reactor for efficient proteolysis
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A nanoporous reactor for efficient proteolysis

机译：用于高效蛋白水解的纳米孔反应器

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摘要

A nanoreactor based on mesoporous silicates is described for efficient tryptic digestion of proteins within the mesochannels. Cyano-functionalized mesoporous silicate (CNS), with an average pore diameter of 18 nm, is a good support for trypsin, with rapid in situ digestion of the model proteins, cytochrome c and myoglobin. The generated peptides were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Proteolysis by trypsin-CNS is much more efficient than in-solution digestion, which can be attributed to nanoscopic confinement and concentration enrichment of the substrate within the mesopores. Proteins at concentrations of 2 ng mu L-1 were successfully identified after digestion for 20 min. A biological complex sample extracted from the cytoplasm of human liver tissue was digested by using the CNS-based reactor. Coupled with reverse-phase HPLC and MALDI-TOF MS/MS, 165 proteins were identified after standard protein data searching. This nanoreactor combines the advantages of short digestion time with retention of enzymatic activity, providing a promising way to advance the development of proteomics.
机译：描述了基于介孔硅酸盐的纳米反应器，用于高效胰蛋白酶消化介孔通道内的蛋白质。氰基官能化的介孔硅酸盐（CNS）平均孔径为18 nm，是对胰蛋白酶的良好支持，可快速原位消化模型蛋白，细胞色素c和肌红蛋白。通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）分析生成的肽。胰蛋白酶-CNS的蛋白水解比溶液中消化的效率要高得多，这可以归因于中孔内底物的纳米级限制和浓度富集。消化20分钟后，成功鉴定出浓度为2 ng mu L-1的蛋白质。使用基于CNS的反应器消化从人肝组织细胞质中提取的生物复合物样品。结合反相HPLC和MALDI-TOF MS / MS，在标准蛋白质数据搜索后鉴定出165种蛋白质。这种纳米反应器结合了消化时间短和酶活性得以保留的优点，为推进蛋白质组学的发展提供了一种有希望的方法。

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