PEGylation and antioxidant effects of a human glutathione peroxidase 1 mutant

Human glutathione peroxidase1 (hGPx1) is a good antioxidant and potential drug, but the limited availability and poor stability of hGPx1 have affected its development and application. To solve this problem, we prepared a hGPx1 mutant (GPx1M) with high activity in an BL21(DE3)cys auxotrophic strain u...

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Published in:Aging (Albany, NY.) NY.), 2022-01, Vol.14 (1), p.443-461
Main Authors: Zhang, Guang-Yuan, Wang, Yan-Wei, Guo, Li-Ying, Lin, Liang-Ru, Niu, Shao-Peng, Xiong, Chang-Hao, Wei, Jing-Yan
Format: Article
Language:English
Subjects:
Animals
Antioxidants - chemistry
Antioxidants - metabolism
Antioxidants - pharmacology
Cell Line
Drug Design
Escherichia coli
Glutathione Peroxidase - chemistry
Glutathione Peroxidase - metabolism
Humans
Hydrogen-Ion Concentration
Models, Molecular
Mutation
Myocytes, Cardiac
Polyethylene Glycols - chemistry
Protein Conformation
Protein Stability
Rats
Rats, Sprague-Dawley
Research Paper
Succinimides - chemistry
Temperature
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Summary:Human glutathione peroxidase1 (hGPx1) is a good antioxidant and potential drug, but the limited availability and poor stability of hGPx1 have affected its development and application. To solve this problem, we prepared a hGPx1 mutant (GPx1M) with high activity in an BL21(DE3)cys auxotrophic strain using a single protein production (SPP) system. In this study, the GPx1M was conjugated with methoxypolyethylene glycol-succinimidyl succinate (SS-mPEG, = 5 kDa) chains to enhance its stability. SS-mPEG-GPx1M and GPx1M exhibited similar enzymatic activity and stability toward pH and temperature change, and in a few cases, SS-mPEG-GPx1M was discovered to widen the range of pH stability and increase the temperature stability. Lys 38 was confirmed as PEGylated site by liquid-mass spectrometry. H9c2 cardiomyoblast cells and Sprague-Dawley (SD) rats were used to evaluate the effects of GPx1M and SS-mPEG-GPx1M on preventing or alleviating adriamycin (ADR)-mediated cardiotoxicity, respectively. The results indicated that GPx1M and SS-mPEG-GPx1M had good antioxidant effects and , and the effect of SS-mPEG-GPx1M is more prominent than GPx1M . Thus, PEGylation might be a promising method for the application of GPx1M as an important antioxidant and potential drug.
ISSN:1945-4589
1945-4589
DOI:10.18632/aging.203822