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Immunologic function and molecular insight of recombinant interleukin-18
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Metadata
Document Title
Immunologic function and molecular insight of recombinant interleukin-18
Author
Saetang J., Puseenam A., Roongsawang N., Voravuthikunchai S.P., Sangkhathat S., Tipmanee V.
Name from Authors Collection
Scopus Author ID
26427756600
Affiliations
Department of Biomedical Sciences, Faculty of Medicine, Prince of Songkla University, Songkhla, 90110, Thailand; Graduate School, Prince of Songkla University, Songkhla, 90110, Thailand; Microbial Cell Factory Laboratory, Bioresources Technology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, Thailand; Department of Microbiology, Natural Product Research Center of Excellence, Faculty of Science, Prince of Songkla University, Songkhla, 90110, Thailand; Department of Surgery, Faculty of Medicine, Prince of Songkla University, Songkhla, 90110, Thailand
Type
Article
Source Title
PLoS ONE
ISSN
19326203
Year
2016
Volume
11
Issue
8
Open Access
Gold, Green
Publisher
Public Library of Science
DOI
10.1371/journal.pone.0160321
Abstract
In recent years, cytokine-mediated therapy has emerged as further advance alternative in cancer therapy. Interleukin-18 (IL-18) has exhibited interesting anti-cancer properties especially when combined with IL-12. We engineered IL-18 in order to improve its activity using single point mutagenesis. IL-18 mutants were constructed according to binding residues and polarity which we tried to increase polarity in M33Q and M60Q, enhanced cationicity in E6K, and flexibility in T63A. All IL-18 proteins were expressed in Pichia pastoris, purified, and then measured the activity by treating with the NK-92MI cell line to evaluate interferon-γ (IFN-γ) stimulation. The E6K and T63A mutant forms showed higher activity with respect to native proteins at the concentration of 200 ng mL-1 by inducing the expression of IFN-γ, about factors of 9 and 4, respectively. Meanwhile, M33Q and M60Q had no significant activity to induce IFN-γ. Interestingly, the combination of E6K and T63A mutations could synergize the induction activity of IL-18 to be 16 times at 200 ng mL-1. Furthermore, molecular dynamics studies have elucidated the effect due to mutation on conformation of the binding site of IL-18. The results turn out that E6K provides structural perseverance against mutation, while M33Q and M60Q promote vivid overall change in protein conformation, especially at the binding site. For T63A, mutation yields small difference in structure but clearly increases structural flexibility. However, a small structural change was observed when T63A was combined with E6K. Our research resulted in a novel version of IL-18 which could be a new key candidate for cytokine-mediated therapy. © 2016 Saetang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding Sponsor
Higher Education Research Promotion; Office of the Higher Education Commission; National Research Council of Thailand; Faculty of Medicine, Prince of Songkla University
License
CC BY
Rights
Author
Publication Source
Scopus