Poly (I:C) and hyaluronic acid directly interact with NLRP3, resulting in the assembly of NLRP3 and ASC in a cell-free system
In the NLR family, pyrin domain containing 3 (NLRP3) is an intracellular pattern recognition receptor that activates pro-caspase-1, leading to IL-1β and IL-18 processing and activation in a large complex called the NLRP3 inflammasome. Since various pathogens or endogenous metabolites have been repor...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
SAGE Publishing
2017-08-01
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| Series: | European Journal of Inflammation |
| Online Access: | https://doi.org/10.1177/1721727X17711047 |
| _version_ | 1818609908175601664 |
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| author | Naoe Kaneko Yuki Ito Tomoyuki Iwasaki Hiroyuki Takeda Tatsuya Sawasaki Kiyoshi Migita Kazunaga Agematsu Tomohiro Koga Atsushi Kawakami Akihiro Yachie Koh-ichiro Yoshiura Shinnosuke Morikawa Mie Kurata Junya Masumoto |
| author_facet | Naoe Kaneko Yuki Ito Tomoyuki Iwasaki Hiroyuki Takeda Tatsuya Sawasaki Kiyoshi Migita Kazunaga Agematsu Tomohiro Koga Atsushi Kawakami Akihiro Yachie Koh-ichiro Yoshiura Shinnosuke Morikawa Mie Kurata Junya Masumoto |
| author_sort | Naoe Kaneko |
| collection | DOAJ |
| description | In the NLR family, pyrin domain containing 3 (NLRP3) is an intracellular pattern recognition receptor that activates pro-caspase-1, leading to IL-1β and IL-18 processing and activation in a large complex called the NLRP3 inflammasome. Since various pathogens or endogenous metabolites have been reported to stimulate NLRP3 inflammasome, the interaction between NLRP3 and ASC induced by these stimulants may be an attractive drug target for NLRP3-related diseases, called inflammasomopathies. However, the endogenous ligand that directly interacts with NLRP3, leading to binding to ASC, remains unclear. Therefore, we developed a cell-free system consisting of NLRP3, ASC, and pro-caspase-1 or ASC and NLRP3 with an amplified luminescent proximity homogeneous assay (ALPHA). ALPHA signals of the interaction between NLRP3 and ASC were not enhanced following an incubation without any ligand, whereas strong ALPHA signals for the interaction between NLRP3 and ASC and between NLRP3 and pro-caspase-1 with the adaptor ASC were observed upon an incubation with poly (I:C) and hyaluronic acid (HA). Poly (I:C) and HA both directly interacted with NLRP3 within a specific concentration. These results suggest that NLRP3 directly interacts with intrinsic RNA and HA, which is followed by the activation of NLRP3 inflammasome, and the cell-free system consisting of NLRP3 and ASC, or NLRP3, ASC, and pro-caspase-1 may be a useful tool for elucidating the pathogenesis of inflammasomopathies and developing target therapeutics. |
| first_indexed | 2024-12-16T15:06:00Z |
| format | Article |
| id | doaj.art-2b4f051e8ddc4634a8aff09e6f57abfb |
| institution | Directory Open Access Journal |
| issn | 1721-727X |
| language | English |
| last_indexed | 2024-12-16T15:06:00Z |
| publishDate | 2017-08-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | European Journal of Inflammation |
| spelling | doaj.art-2b4f051e8ddc4634a8aff09e6f57abfb2022-12-21T22:27:08ZengSAGE PublishingEuropean Journal of Inflammation1721-727X2017-08-011510.1177/1721727X17711047Poly (I:C) and hyaluronic acid directly interact with NLRP3, resulting in the assembly of NLRP3 and ASC in a cell-free systemNaoe Kaneko0Yuki Ito1Tomoyuki Iwasaki2Hiroyuki Takeda3Tatsuya Sawasaki4Kiyoshi Migita5Kazunaga Agematsu6Tomohiro Koga7Atsushi Kawakami8Akihiro Yachie9Koh-ichiro Yoshiura10Shinnosuke Morikawa11Mie Kurata12Junya Masumoto13Department of Pathology, Proteo-Science Center and Graduate School of Medicine, Ehime University, Toon, JapanDepartment of Pathology, Proteo-Science Center and Graduate School of Medicine, Ehime University, Toon, JapanDepartment of Pathology, Proteo-Science Center and Graduate School of Medicine, Ehime University, Toon, JapanDivision of Cell-Free Sciences, Proteo-Science Center, Ehime University, Matsuyama, JapanDivision of Cell-Free Sciences, Proteo-Science Center, Ehime University, Matsuyama, JapanDepartment of Gastroenterology and Rheumatology, School of Medicine, Fukushima Medical University, Fukushima, JapanDepartment of Infectious Immunology, Graduate School of Medicine, Shinshu University, Matsumoto, JapanMedical Education Development Center, Nagasaki University Hospital, Nagasaki, JapanUnit of Translational Medicine, Department of Immunology & Rheumatology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, JapanDepartment of Pediatrics, School of Medicine, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kanazawa, JapanDepartment of Human Genetics, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, JapanDepartment of Pathology, Proteo-Science Center and Graduate School of Medicine, Ehime University, Toon, JapanDepartment of Pathology, Proteo-Science Center and Graduate School of Medicine, Ehime University, Toon, JapanDepartment of Pathology, Proteo-Science Center and Graduate School of Medicine, Ehime University, Toon, JapanIn the NLR family, pyrin domain containing 3 (NLRP3) is an intracellular pattern recognition receptor that activates pro-caspase-1, leading to IL-1β and IL-18 processing and activation in a large complex called the NLRP3 inflammasome. Since various pathogens or endogenous metabolites have been reported to stimulate NLRP3 inflammasome, the interaction between NLRP3 and ASC induced by these stimulants may be an attractive drug target for NLRP3-related diseases, called inflammasomopathies. However, the endogenous ligand that directly interacts with NLRP3, leading to binding to ASC, remains unclear. Therefore, we developed a cell-free system consisting of NLRP3, ASC, and pro-caspase-1 or ASC and NLRP3 with an amplified luminescent proximity homogeneous assay (ALPHA). ALPHA signals of the interaction between NLRP3 and ASC were not enhanced following an incubation without any ligand, whereas strong ALPHA signals for the interaction between NLRP3 and ASC and between NLRP3 and pro-caspase-1 with the adaptor ASC were observed upon an incubation with poly (I:C) and hyaluronic acid (HA). Poly (I:C) and HA both directly interacted with NLRP3 within a specific concentration. These results suggest that NLRP3 directly interacts with intrinsic RNA and HA, which is followed by the activation of NLRP3 inflammasome, and the cell-free system consisting of NLRP3 and ASC, or NLRP3, ASC, and pro-caspase-1 may be a useful tool for elucidating the pathogenesis of inflammasomopathies and developing target therapeutics.https://doi.org/10.1177/1721727X17711047 |
| spellingShingle | Naoe Kaneko Yuki Ito Tomoyuki Iwasaki Hiroyuki Takeda Tatsuya Sawasaki Kiyoshi Migita Kazunaga Agematsu Tomohiro Koga Atsushi Kawakami Akihiro Yachie Koh-ichiro Yoshiura Shinnosuke Morikawa Mie Kurata Junya Masumoto Poly (I:C) and hyaluronic acid directly interact with NLRP3, resulting in the assembly of NLRP3 and ASC in a cell-free system European Journal of Inflammation |
| title | Poly (I:C) and hyaluronic acid directly interact with NLRP3, resulting in the assembly of NLRP3 and ASC in a cell-free system |
| title_full | Poly (I:C) and hyaluronic acid directly interact with NLRP3, resulting in the assembly of NLRP3 and ASC in a cell-free system |
| title_fullStr | Poly (I:C) and hyaluronic acid directly interact with NLRP3, resulting in the assembly of NLRP3 and ASC in a cell-free system |
| title_full_unstemmed | Poly (I:C) and hyaluronic acid directly interact with NLRP3, resulting in the assembly of NLRP3 and ASC in a cell-free system |
| title_short | Poly (I:C) and hyaluronic acid directly interact with NLRP3, resulting in the assembly of NLRP3 and ASC in a cell-free system |
| title_sort | poly i c and hyaluronic acid directly interact with nlrp3 resulting in the assembly of nlrp3 and asc in a cell free system |
| url | https://doi.org/10.1177/1721727X17711047 |
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