Effects of Glutathionylation on Guanylyltransferase Activity of NS5 N-terminal Capping Domain from Dengue, Japanese Encephalitis, and Zika Viruses
Issued Date
2023-01-01
Resource Type
ISSN
09298665
eISSN
18755305
Scopus ID
2-s2.0-85164208418
Pubmed ID
37076471
Journal Title
Protein and Peptide Letters
Volume
30
Issue
5
Start Page
439
End Page
447
Rights Holder(s)
SCOPUS
Bibliographic Citation
Protein and Peptide Letters Vol.30 No.5 (2023) , 439-447
Suggested Citation
Saisawang C., Reamtong O., Nachampa I., Petcharat P., Priewkhiew S., Sakdee S., Wongsantichon J., Ketterman A.J. Effects of Glutathionylation on Guanylyltransferase Activity of NS5 N-terminal Capping Domain from Dengue, Japanese Encephalitis, and Zika Viruses. Protein and Peptide Letters Vol.30 No.5 (2023) , 439-447. 447. doi:10.2174/0929866530666230418101606 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/87952
Title
Effects of Glutathionylation on Guanylyltransferase Activity of NS5 N-terminal Capping Domain from Dengue, Japanese Encephalitis, and Zika Viruses
Other Contributor(s)
Abstract
Background: Glutathionylation is a protein post-translational modification triggered by oxidative stress. The susceptible proteins are modified by the addition of glutathione to specific cysteine residues. Virus infection also induces oxidative stress in the cell, which affects cellular homeostasis. It is not just the cellular proteins but the viral proteins that can also be modified by glutathionylation events, thereby impacting the function of the viral proteins. Objectives: This study was conducted to identify the effects of modification by glutathionylation on the guanylyltransferase activity of NS5 and identify the cysteine residues modified for the three flavivirus NS5 proteins. Methods: The capping domain of NS5 proteins from 3 flaviviruses was cloned and expressed as recombinant proteins. A gel-based assay for guanylyltransferase activity was performed using a GTP analog labeled with the fluorescent dye Cy5 as substrate. The protein modification by glutathionylation was induced by GSSG and evaluated by western blot. The reactive cysteine residues were identified by mass spectrometry. Results: It was found that the three flavivirus proteins behaved in a similar fashion with increasing glutathionylation yielding decreased guanylyltransferase activity. The three proteins also possessed conserved cysteines and they appeared to be modified for all three proteins. Conclusion: The glutathionylation appeared to induce conformational changes that affect enzyme activity. The conformational changes might also create binding sites for host cell protein interactions at later stages of viral propagation with the glutathionylation event, thereby serving as a switch for function change.