Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like Activities
Zhuofu Wu, 1, 2 Sitong Zhang, 1 Xiaojun Wang, 2 Can Cai, 2 Guang Chen, 1 Li Ma 2 1Key Laboratory of Straw Biology and Utilization, The Ministry of Education, College of Life Science, Jilin Agricultural University, Changchun 130118, People’s Republic of China; 2Department of Physics, Geor...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Dove Medical Press
2020-01-01
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| Series: | International Journal of Nanomedicine |
| Subjects: | |
| Online Access: | https://www.dovepress.com/nitroxide-modified-protein-incorporated-nanoflowers-with-dual-enzyme-l-peer-reviewed-article-IJN |
| _version_ | 1818998954676715520 |
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| author | Wu Z Zhang S Wang X Cai C Chen G Ma L |
| author_facet | Wu Z Zhang S Wang X Cai C Chen G Ma L |
| author_sort | Wu Z |
| collection | DOAJ |
| description | Zhuofu Wu, 1, 2 Sitong Zhang, 1 Xiaojun Wang, 2 Can Cai, 2 Guang Chen, 1 Li Ma 2 1Key Laboratory of Straw Biology and Utilization, The Ministry of Education, College of Life Science, Jilin Agricultural University, Changchun 130118, People’s Republic of China; 2Department of Physics, Georgia Southern University, Statesboro, GA 30460, USACorrespondence: Guang ChenCollege of Life Science, Jilin Agricultural University, No. 2888 Xincheng Avenue, Changchun 130118, People’s Republic of ChinaTel/Fax +86 431 8453 2942Email guangc61@gmail.com
Li MaDepartment of Physics, Georgia Southern University, 65 Georgia Avenue, Math/Physics Building Room 1003, Statesboro, GA 30460, USATel +1 912 478 5950Fax +1 912 478 0471Email lma@GeorgiaSouthern.eduPurpose: 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 2O 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 2O 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 2O 2 not only obey Michaelis–Menton kinetics, but that the nanoflowers also possess a higher affinity for H 2O 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.Keywords: catalase, superoxide dismutase, polynitroxylated human serum albumin, reactive oxygen species, copper-based nanoflowers |
| first_indexed | 2024-12-20T22:09:44Z |
| format | Article |
| id | doaj.art-537f764a351445469cc5f5e02cebeb38 |
| institution | Directory Open Access Journal |
| issn | 1178-2013 |
| language | English |
| last_indexed | 2024-12-20T22:09:44Z |
| publishDate | 2020-01-01 |
| publisher | Dove Medical Press |
| record_format | Article |
| series | International Journal of Nanomedicine |
| spelling | doaj.art-537f764a351445469cc5f5e02cebeb382022-12-21T19:25:12ZengDove Medical PressInternational Journal of Nanomedicine1178-20132020-01-01Volume 1526327351144Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like ActivitiesWu ZZhang SWang XCai CChen GMa LZhuofu Wu, 1, 2 Sitong Zhang, 1 Xiaojun Wang, 2 Can Cai, 2 Guang Chen, 1 Li Ma 2 1Key Laboratory of Straw Biology and Utilization, The Ministry of Education, College of Life Science, Jilin Agricultural University, Changchun 130118, People’s Republic of China; 2Department of Physics, Georgia Southern University, Statesboro, GA 30460, USACorrespondence: Guang ChenCollege of Life Science, Jilin Agricultural University, No. 2888 Xincheng Avenue, Changchun 130118, People’s Republic of ChinaTel/Fax +86 431 8453 2942Email guangc61@gmail.com Li MaDepartment of Physics, Georgia Southern University, 65 Georgia Avenue, Math/Physics Building Room 1003, Statesboro, GA 30460, USATel +1 912 478 5950Fax +1 912 478 0471Email lma@GeorgiaSouthern.eduPurpose: 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 2O 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 2O 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 2O 2 not only obey Michaelis–Menton kinetics, but that the nanoflowers also possess a higher affinity for H 2O 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.Keywords: catalase, superoxide dismutase, polynitroxylated human serum albumin, reactive oxygen species, copper-based nanoflowershttps://www.dovepress.com/nitroxide-modified-protein-incorporated-nanoflowers-with-dual-enzyme-l-peer-reviewed-article-IJNcatalasesuperoxide dismutasepolynitroxylated human serum albuminreactive oxygen species |
| spellingShingle | Wu Z Zhang S Wang X Cai C Chen G Ma L Nitroxide-Modified Protein-Incorporated Nanoflowers with Dual Enzyme-Like Activities International Journal of Nanomedicine catalase superoxide dismutase polynitroxylated human serum albumin reactive oxygen species |
| 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 | catalase superoxide dismutase polynitroxylated human serum albumin reactive oxygen species |
| url | https://www.dovepress.com/nitroxide-modified-protein-incorporated-nanoflowers-with-dual-enzyme-l-peer-reviewed-article-IJN |
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