PMID- 27483370
OWN - NLM
STAT- MEDLINE
DCOM- 20170726
LR  - 20181202
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 11
IP  - 8
DP  - 2016
TI  - Immunologic Function and Molecular Insight of Recombinant Interleukin-18.
PG  - e0160321
LID - 10.1371/journal.pone.0160321 [doi]
LID - e0160321
AB  - 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-gamma (IFN-gamma) 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-gamma, about factors of 9 and 4, respectively. 
      Meanwhile, M33Q and M60Q had no significant activity to induce IFN-gamma. 
      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.
FAU - Saetang, Jirakrit
AU  - Saetang J
AD  - Department of Biomedical Sciences, Faculty of Medicine, Prince of Songkla 
      University, Songkhla, 90110, Thailand.
AD  - Graduate School, Prince of Songkla University, Songkhla, 90110, Thailand.
FAU - Puseenam, Aekkachai
AU  - Puseenam A
AD  - 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.
FAU - Roongsawang, Niran
AU  - Roongsawang N
AD  - 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.
FAU - Voravuthikunchai, Supayang Piyawan
AU  - Voravuthikunchai SP
AD  - Department of Microbiology and Natural Product Research Center of Excellence, 
      Faculty of Science, Prince of Songkla University, Songkhla, 90110, Thailand.
FAU - Sangkhathat, Surasak
AU  - Sangkhathat S
AD  - Department of Biomedical Sciences, Faculty of Medicine, Prince of Songkla 
      University, Songkhla, 90110, Thailand.
AD  - Department of Surgery, Faculty of Medicine, Prince of Songkla University, 
      Songkhla, 90110, Thailand.
FAU - Tipmanee, Varomyalin
AU  - Tipmanee V
AUID- ORCID: 0000-0001-6017-7519
AD  - Department of Biomedical Sciences, Faculty of Medicine, Prince of Songkla 
      University, Songkhla, 90110, Thailand.
LA  - eng
PT  - Journal Article
DEP - 20160802
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Interleukin-18)
RN  - 0 (Receptors, Interleukin-18)
RN  - 0 (Recombinant Proteins)
RN  - 82115-62-6 (Interferon-gamma)
SB  - IM
MH  - Amino Acid Sequence
MH  - Binding Sites
MH  - Cell Line, Tumor
MH  - Cloning, Molecular
MH  - Gene Expression
MH  - Humans
MH  - Interferon-gamma/*biosynthesis/metabolism
MH  - Interleukin-18/*chemistry/genetics/immunology/pharmacology
MH  - Killer Cells, Natural/cytology/*drug effects/immunology
MH  - Kinetics
MH  - Lymphocyte Activation/*drug effects
MH  - Models, Molecular
MH  - Molecular Weight
MH  - Pichia/genetics/metabolism
MH  - Protein Binding
MH  - Protein Conformation, alpha-Helical
MH  - Protein Conformation, beta-Strand
MH  - Protein Engineering
MH  - Protein Interaction Domains and Motifs
MH  - Receptors, Interleukin-18/*chemistry/genetics/immunology
MH  - Recombinant Proteins/chemistry/genetics/immunology
MH  - Sequence Alignment
MH  - Structure-Activity Relationship
MH  - Substrate Specificity
PMC - PMC4970725
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2016/08/03 06:00
MHDA- 2017/07/27 06:00
PMCR- 2016/08/02
CRDT- 2016/08/03 06:00
PHST- 2016/02/01 00:00 [received]
PHST- 2016/07/18 00:00 [accepted]
PHST- 2016/08/03 06:00 [entrez]
PHST- 2016/08/03 06:00 [pubmed]
PHST- 2017/07/27 06:00 [medline]
PHST- 2016/08/02 00:00 [pmc-release]
AID - PONE-D-16-02822 [pii]
AID - 10.1371/journal.pone.0160321 [doi]
PST - epublish
SO  - PLoS One. 2016 Aug 2;11(8):e0160321. doi: 10.1371/journal.pone.0160321. 
      eCollection 2016.