Functionalization of Mesoporous Silica with a G-A-Mismatched dsDNA Chain for Efficient Identification and Selective Capturing of the MutY Protein,ACS Applied Materials & Interfaces

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Functionalization of Mesoporous Silica with a G-A-Mismatched dsDNA Chain for Efficient Identification and Selective Capturing of the MutY Protein
ACS Applied Materials & Interfaces ( IF 8.3 ) Pub Date : 2023-02-09 , DOI: 10.1021/acsami.2c19257
Jingqi Sun ₁ , Jianhua Wang ₁ , Xuwei Chen ₁

Affiliation

MUTYH adenine DNA glycosylase and its homologous protein (collectively MutY) are typical DNA glycosylases with a [4Fe4S] cluster and a helix–hairpin–helix (HhH) motif in its structure. In the present work, the binding behaviors of the MutY protein to dsDNA containing different base mismatches were investigated. The type and distribution of base mismatch in the dsDNA chain were found to influence the DNA–protein binding interaction greatly. The [4Fe4S] cluster of the MutY protein is able to identify a G-A mismatch in the dsDNA chain specifically by monitoring the anomalies of charge transport in the dsDNA chain, allowing the entrance of the identified dsDNA chain into the internal cavity of the MutY protein and the strong DNA–protein binding at the HhH motif of the protein through multiple H-bonds. The dsDNA chain with a centrally located G-A mismatch is thus functionalized on mesoporous silica (MSN) via amination reaction, and the obtained dsDNA(G-A)@MSN is used as a powerful sorbent for the selective capturing of the MutY protein from complex samples. By using 0.5% NH₃·H₂O (m/v) as a stripping reagent, efficient isolation of the MutY protein from different cell lines and bacteria is achieved and the recovered MutY protein is demonstrated to maintain favorable DNA adenine glycosylase activity.

中文翻译：

使用 GA 不匹配的 dsDNA 链对介孔二氧化硅进行功能化，以有效识别和选择性捕获 MutY 蛋白

MUTYH 腺嘌呤 DNA 糖基化酶及其同源蛋白（统称为 MutY）是典型的 DNA 糖基化酶，其结构中具有 [4Fe4S] 簇和螺旋-发夹-螺旋 (HhH) 基序。在本工作中，研究了 MutY 蛋白与含有不同碱基错配的 dsDNA 的结合行为。研究发现双链 DNA 链中碱基错配的类型和分布对 DNA-蛋白质结合相互作用有很大影响。 MutY 蛋白的 [4Fe4S] 簇能够通过监测 dsDNA 链中电荷传输的异常来特异性识别 dsDNA 链中的 GA 错配，从而允许所识别的 dsDNA 链进入 MutY 蛋白的内腔并DNA-蛋白质通过多个氢键在蛋白质的 HhH 基序上强结合。因此，具有中心 GA 错配的 dsDNA 链通过胺化反应在介孔二氧化硅 (MSN) 上功能化，所得 dsDNA(GA)@MSN 用作强大的吸附剂，用于从复杂样品中选择性捕获 MutY 蛋白。通过使用0.5% NH ₃ ·H ₂ O (m/v)作为剥离试剂，实现了从不同细胞系和细菌中有效分离MutY蛋白，并证明回收的MutY蛋白保持良好的DNA腺嘌呤糖基化酶活性。

更新日期：2023-02-09

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