Cargando…
Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like Activities
PURPOSE: Combined superoxide dismutase (SOD)/catalase mimetics have attracted much attention because of their efficacy against reactive oxygen species-associated diseases; however, their application is often limited owing to their poor stability and the absence of favorable grafting sites. To addres...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Dove
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6970245/ https://www.ncbi.nlm.nih.gov/pubmed/32021179 http://dx.doi.org/10.2147/IJN.S220718 |
_version_ | 1783489479565115392 |
---|---|
author | Wu, Zhuofu Zhang, Sitong Wang, Xiaojun Cai, Can Chen, Guang Ma, Li |
author_facet | Wu, Zhuofu Zhang, Sitong Wang, Xiaojun Cai, Can Chen, Guang Ma, Li |
author_sort | Wu, Zhuofu |
collection | PubMed |
description | PURPOSE: Combined superoxide dismutase (SOD)/catalase mimetics have attracted much attention because of their efficacy against reactive oxygen species-associated diseases; however, their application is often limited owing to their poor stability and the absence of favorable grafting sites. To address this, we developed a new class of SOD/catalase mimetics based on hybrid nanoflowers, which exhibit superior stability and possess the desired grafting sites for drugs and endogenous molecules. METHODS: In this work, for the first time, we used polynitroxylated human serum albumin (PNA) to mediate the formation of hybrid copper-based nanoflowers. H(2)O(2) depletion and O(2) evolution assays were first performed to determine the catalase-like activity of the hybrid nanoflowers. Next, the xanthine oxidase/cytochrome c method was used to assay the SOD-like activity of the nanoflowers. Further characteristics of the nanoflowers were evaluated using scanning electron microscopy (SEM), electron paramagnetic resonance (EPR), and Fourier-transform infrared spectroscopy (FTIR). Operational stability was assessed via the reusability assay. RESULTS: The H(2)O(2) depletion and O(2) evolution assays indicated that PNA-incorporated nanoflowers have genuine catalase-like activity. Kinetic analysis revealed that the reactions of the incorporated nanoflowers with H(2)O(2) not only obey Michaelis–Menton kinetics, but that the nanoflowers also possess a higher affinity for H(2)O(2) than that of native catalase. The FTIR spectra corroborated the presence of PNA in the hybrid nanoflowers, while the EPR spectra confirmed the intermolecular interaction of nitroxides bound to the human serum albumin incorporated into the nanoflowers. The remarkable operational reproducibility of the hybrid nanoflowers in catalase-like and SOD-like reactions was verified across successive batches. CONCLUSION: Herein, a comparison of Michaelis constants showed that the hybrid nanoflower, a catalase mimetics, outperforms the native catalase. Acting as a “better-than-nature” enzyme mimetics, the hybrid nanoflower with superior stability and desired ligand grafting sites will find widespread utilization in the medical sciences. |
format | Online Article Text |
id | pubmed-6970245 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Dove |
record_format | MEDLINE/PubMed |
spelling | pubmed-69702452020-02-04 Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like Activities Wu, Zhuofu Zhang, Sitong Wang, Xiaojun Cai, Can Chen, Guang Ma, Li Int J Nanomedicine Original Research PURPOSE: Combined superoxide dismutase (SOD)/catalase mimetics have attracted much attention because of their efficacy against reactive oxygen species-associated diseases; however, their application is often limited owing to their poor stability and the absence of favorable grafting sites. To address this, we developed a new class of SOD/catalase mimetics based on hybrid nanoflowers, which exhibit superior stability and possess the desired grafting sites for drugs and endogenous molecules. METHODS: In this work, for the first time, we used polynitroxylated human serum albumin (PNA) to mediate the formation of hybrid copper-based nanoflowers. H(2)O(2) depletion and O(2) evolution assays were first performed to determine the catalase-like activity of the hybrid nanoflowers. Next, the xanthine oxidase/cytochrome c method was used to assay the SOD-like activity of the nanoflowers. Further characteristics of the nanoflowers were evaluated using scanning electron microscopy (SEM), electron paramagnetic resonance (EPR), and Fourier-transform infrared spectroscopy (FTIR). Operational stability was assessed via the reusability assay. RESULTS: The H(2)O(2) depletion and O(2) evolution assays indicated that PNA-incorporated nanoflowers have genuine catalase-like activity. Kinetic analysis revealed that the reactions of the incorporated nanoflowers with H(2)O(2) not only obey Michaelis–Menton kinetics, but that the nanoflowers also possess a higher affinity for H(2)O(2) than that of native catalase. The FTIR spectra corroborated the presence of PNA in the hybrid nanoflowers, while the EPR spectra confirmed the intermolecular interaction of nitroxides bound to the human serum albumin incorporated into the nanoflowers. The remarkable operational reproducibility of the hybrid nanoflowers in catalase-like and SOD-like reactions was verified across successive batches. CONCLUSION: Herein, a comparison of Michaelis constants showed that the hybrid nanoflower, a catalase mimetics, outperforms the native catalase. Acting as a “better-than-nature” enzyme mimetics, the hybrid nanoflower with superior stability and desired ligand grafting sites will find widespread utilization in the medical sciences. Dove 2020-01-15 /pmc/articles/PMC6970245/ /pubmed/32021179 http://dx.doi.org/10.2147/IJN.S220718 Text en © 2020 Wu et al. http://creativecommons.org/licenses/by-nc/3.0/ This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php). |
spellingShingle | Original Research Wu, Zhuofu Zhang, Sitong Wang, Xiaojun Cai, Can Chen, Guang Ma, Li Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like Activities |
title | Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like Activities |
title_full | Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like Activities |
title_fullStr | Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like Activities |
title_full_unstemmed | Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like Activities |
title_short | Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like Activities |
title_sort | nitroxide-modified protein-incorporated nanoflowers with dual enzyme-like activities |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6970245/ https://www.ncbi.nlm.nih.gov/pubmed/32021179 http://dx.doi.org/10.2147/IJN.S220718 |
work_keys_str_mv | AT wuzhuofu nitroxidemodifiedproteinincorporatednanoflowerswithdualenzymelikeactivities AT zhangsitong nitroxidemodifiedproteinincorporatednanoflowerswithdualenzymelikeactivities AT wangxiaojun nitroxidemodifiedproteinincorporatednanoflowerswithdualenzymelikeactivities AT caican nitroxidemodifiedproteinincorporatednanoflowerswithdualenzymelikeactivities AT chenguang nitroxidemodifiedproteinincorporatednanoflowerswithdualenzymelikeactivities AT mali nitroxidemodifiedproteinincorporatednanoflowerswithdualenzymelikeactivities |