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一种用于检测源自HeLa细胞线粒体释放的超氧阴离子的羟基自由基的酶包封MOF@MOF纳米复合材料。

An Enzyme-Encapsulated MOF@MOF Nanocomposite for Detecting HO Derived From Superoxide Anion Released by Mitochondria of HeLa Cells.

作者信息

Niu Jiaqi, Jin Xiaoxin, Wang Xingqi, Ren Zhenhua, Li Bingjie, Liu Xiaoqiang, Wong Danny K Y

机构信息

Henan International Joint Laboratory of Medicinal Plants Utilization, College of Chemistry and Molecular Sciences, Henan University, Zhengzhou, Henan, 450046, P. R. China.

Department of Applied BioSciences, Macquarie University, Sydney, NSW, 2109, Australia.

出版信息

Small Methods. 2025 Mar;9(3):e2401070. doi: 10.1002/smtd.202401070. Epub 2024 Sep 16.

Abstract

In this work, a horseradish peroxidase (HRP)-encapsulated metal organic framework (MOF)@MOF nanocomposite is developed for detecting HO converted by dismutation of superoxide anions released from live HeLa mitochondria. Initially, an HRP-polyacrylic acid cluster is incorporated on a mesoporous, peroxidase-like Cu/Co-1,4-benzenedicarboxylate (BDC) MOF platform to avoid structural change and deactivation of HRP through its interactions with MOF metal ions. Additionally, a Cu/Co-BDC(HRP)@1,3,5-benzenetricarboxyate (BTC) core-shell MOF/MOF structure, also with peroxide-like properties, serves as a protective matrix for HRP. Then, ultrathin porous carbon shells (UPCS) are adopted to improve the electrical conductivity of the MOF@MOF. The Cu/Co-BDC(HRP)@BTC|UPCS sensing platform exhibits two linear ranges of 0.05-1 µM and 1-1000 µM with a sensitivity of 172 mA mM cm and 1.63 mA mM cm, respectively. A limit of detection of 0.057 µM, good selectivity and stability over 35 days for HO detection are also achieved. After treating the mitochondrial complex with specific inhibitors, amperometric results at the sensing platform confirmed complex I and III within mitochondria as the main electron leakage sites in the electron transfer chains. Therefore, this sensing platform provides a tool that may aid in predicting and even developing treatments for some oxidative stress diseases caused by mitochondrial abnormalities.

摘要

在这项工作中,开发了一种辣根过氧化物酶(HRP)封装的金属有机框架(MOF)@MOF纳米复合材料,用于检测由活HeLa线粒体释放的超氧阴离子歧化转化产生的HO。最初,将HRP-聚丙烯酸簇掺入介孔的、具有过氧化物酶样性质的Cu/Co-1,4-苯二甲酸(BDC)MOF平台上,以避免HRP通过与MOF金属离子相互作用而发生结构变化和失活。此外,具有类似过氧化物性质的Cu/Co-BDC(HRP)@1,3,5-苯三甲酸(BTC)核壳MOF/MOF结构用作HRP的保护基质。然后,采用超薄多孔碳壳(UPCS)来提高MOF@MOF的电导率。Cu/Co-BDC(HRP)@BTC|UPCS传感平台表现出两个线性范围,分别为0.05 - 1 µM和1 - 1000 µM,灵敏度分别为172 mA mM cm和1.63 mA mM cm。还实现了0.057 µM的检测限、良好的选择性以及在35天内对HO检测的稳定性。在用特定抑制剂处理线粒体复合物后,传感平台上的安培法结果证实线粒体中的复合物I和III是电子传递链中的主要电子泄漏位点。因此,该传感平台提供了一种工具,可能有助于预测甚至开发针对某些由线粒体异常引起的氧化应激疾病的治疗方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/741d/11926491/858cf11938e5/SMTD-9-2401070-g006.jpg

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