Modeling-Enabled Characterization of Novel NLRX1 Ligands.
Nucleotide-binding domain and leucine-rich repeat containing (NLR) family are intracellular sentinels of cytosolic homeostasis that orchestrate immune and inflammatory responses in infectious and immune-mediated diseases. NLRX1 is a mitochondrial-associated NOD-like receptor involved in the modulati...
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4694766?pdf=render |
| _version_ | 1818985432274173952 |
|---|---|
| author | Pinyi Lu Raquel Hontecillas Vida Abedi Shiv Kale Andrew Leber Chase Heltzel Mark Langowski Victoria Godfrey Casandra Philipson Nuria Tubau-Juni Adria Carbo Stephen Girardin Aykut Uren Josep Bassaganya-Riera |
| author_facet | Pinyi Lu Raquel Hontecillas Vida Abedi Shiv Kale Andrew Leber Chase Heltzel Mark Langowski Victoria Godfrey Casandra Philipson Nuria Tubau-Juni Adria Carbo Stephen Girardin Aykut Uren Josep Bassaganya-Riera |
| author_sort | Pinyi Lu |
| collection | DOAJ |
| description | Nucleotide-binding domain and leucine-rich repeat containing (NLR) family are intracellular sentinels of cytosolic homeostasis that orchestrate immune and inflammatory responses in infectious and immune-mediated diseases. NLRX1 is a mitochondrial-associated NOD-like receptor involved in the modulation of immune and metabolic responses. This study utilizes molecular docking approaches to investigate the structure of NLRX1 and experimentally assesses binding to naturally occurring compounds from several natural product and lipid databases. Screening of compound libraries predicts targeting of NLRX1 by conjugated trienes, polyketides, prenol lipids, sterol lipids, and coenzyme A-containing fatty acids for activating the NLRX1 pathway. The ligands of NLRX1 were identified by docking punicic acid (PUA), eleostearic acid (ESA), and docosahexaenoic acid (DHA) to the C-terminal fragment of the human NLRX1 (cNLRX1). Their binding and that of positive control RNA to cNLRX1 were experimentally determined by surface plasmon resonance (SPR) spectroscopy. In addition, the ligand binding sites of cNLRX1 were predicted in silico and validated experimentally. Target mutagenesis studies demonstrate that mutation of 4 critical residues ASP677, PHE680, PHE681, and GLU684 to alanine resulted in diminished affinity of PUA, ESA, and DHA to NLRX1. Consistent with the regulatory actions of NLRX1 on the NF-κB pathway, treatment of bone marrow derived macrophages (BMDM)s with PUA and DHA suppressed NF-κB activity in a NLRX1 dependent mechanism. In addition, a series of pre-clinical efficacy studies were performed using a mouse model of dextran sodium sulfate (DSS)-induced colitis. Our findings showed that the regulatory function of PUA on colitis is NLRX1 dependent. Thus, we identified novel small molecules that bind to NLRX1 and exert anti-inflammatory actions. |
| first_indexed | 2024-12-20T18:34:48Z |
| format | Article |
| id | doaj.art-8c0dafe89c8443a7928c4749099a6093 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-20T18:34:48Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-8c0dafe89c8443a7928c4749099a60932022-12-21T19:29:58ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011012e014542010.1371/journal.pone.0145420Modeling-Enabled Characterization of Novel NLRX1 Ligands.Pinyi LuRaquel HontecillasVida AbediShiv KaleAndrew LeberChase HeltzelMark LangowskiVictoria GodfreyCasandra PhilipsonNuria Tubau-JuniAdria CarboStephen GirardinAykut UrenJosep Bassaganya-RieraNucleotide-binding domain and leucine-rich repeat containing (NLR) family are intracellular sentinels of cytosolic homeostasis that orchestrate immune and inflammatory responses in infectious and immune-mediated diseases. NLRX1 is a mitochondrial-associated NOD-like receptor involved in the modulation of immune and metabolic responses. This study utilizes molecular docking approaches to investigate the structure of NLRX1 and experimentally assesses binding to naturally occurring compounds from several natural product and lipid databases. Screening of compound libraries predicts targeting of NLRX1 by conjugated trienes, polyketides, prenol lipids, sterol lipids, and coenzyme A-containing fatty acids for activating the NLRX1 pathway. The ligands of NLRX1 were identified by docking punicic acid (PUA), eleostearic acid (ESA), and docosahexaenoic acid (DHA) to the C-terminal fragment of the human NLRX1 (cNLRX1). Their binding and that of positive control RNA to cNLRX1 were experimentally determined by surface plasmon resonance (SPR) spectroscopy. In addition, the ligand binding sites of cNLRX1 were predicted in silico and validated experimentally. Target mutagenesis studies demonstrate that mutation of 4 critical residues ASP677, PHE680, PHE681, and GLU684 to alanine resulted in diminished affinity of PUA, ESA, and DHA to NLRX1. Consistent with the regulatory actions of NLRX1 on the NF-κB pathway, treatment of bone marrow derived macrophages (BMDM)s with PUA and DHA suppressed NF-κB activity in a NLRX1 dependent mechanism. In addition, a series of pre-clinical efficacy studies were performed using a mouse model of dextran sodium sulfate (DSS)-induced colitis. Our findings showed that the regulatory function of PUA on colitis is NLRX1 dependent. Thus, we identified novel small molecules that bind to NLRX1 and exert anti-inflammatory actions.http://europepmc.org/articles/PMC4694766?pdf=render |
| spellingShingle | Pinyi Lu Raquel Hontecillas Vida Abedi Shiv Kale Andrew Leber Chase Heltzel Mark Langowski Victoria Godfrey Casandra Philipson Nuria Tubau-Juni Adria Carbo Stephen Girardin Aykut Uren Josep Bassaganya-Riera Modeling-Enabled Characterization of Novel NLRX1 Ligands. PLoS ONE |
| title | Modeling-Enabled Characterization of Novel NLRX1 Ligands. |
| title_full | Modeling-Enabled Characterization of Novel NLRX1 Ligands. |
| title_fullStr | Modeling-Enabled Characterization of Novel NLRX1 Ligands. |
| title_full_unstemmed | Modeling-Enabled Characterization of Novel NLRX1 Ligands. |
| title_short | Modeling-Enabled Characterization of Novel NLRX1 Ligands. |
| title_sort | modeling enabled characterization of novel nlrx1 ligands |
| url | http://europepmc.org/articles/PMC4694766?pdf=render |
| work_keys_str_mv | AT pinyilu modelingenabledcharacterizationofnovelnlrx1ligands AT raquelhontecillas modelingenabledcharacterizationofnovelnlrx1ligands AT vidaabedi modelingenabledcharacterizationofnovelnlrx1ligands AT shivkale modelingenabledcharacterizationofnovelnlrx1ligands AT andrewleber modelingenabledcharacterizationofnovelnlrx1ligands AT chaseheltzel modelingenabledcharacterizationofnovelnlrx1ligands AT marklangowski modelingenabledcharacterizationofnovelnlrx1ligands AT victoriagodfrey modelingenabledcharacterizationofnovelnlrx1ligands AT casandraphilipson modelingenabledcharacterizationofnovelnlrx1ligands AT nuriatubaujuni modelingenabledcharacterizationofnovelnlrx1ligands AT adriacarbo modelingenabledcharacterizationofnovelnlrx1ligands AT stephengirardin modelingenabledcharacterizationofnovelnlrx1ligands AT aykuturen modelingenabledcharacterizationofnovelnlrx1ligands AT josepbassaganyariera modelingenabledcharacterizationofnovelnlrx1ligands |