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Molecular analysis of Fasciola gigantica nuclear receptor subfamily 1-ligand binding domain |
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| รหัสดีโอไอ | |
| Title | Molecular analysis of Fasciola gigantica nuclear receptor subfamily 1-ligand binding domain |
| Creator | Kanokwan Kankul |
| Contributor | Pongsakorn Martviset, Advisor |
| Publisher | Thammasat University |
| Publication Year | 2568 |
| Keyword | Fasciola gigantica, FgNR1-LBD, Bile salts, Molecular docking |
| Abstract | Fasciolosis, also known as liver rot disease, is caused by Fasciola spp., which includes two major species, F. hepatica and F. gigantica. It poses a significant threat to public health and the global economy, affecting both livestock and humans, and causing severe infections in both intrahepatic and extrahepatic areas. Therefore, effective control requires the discovery of new drug targets. In this study, we focus on F. gigantica, which is prevalent in Thailand. Initially, we looked up the habitat of the adult worm. The adult F. gigantica lives in bile ducts, which are rich in bile salts, the molecules that are structurally similar to steroid hormones. Our research group had first identified the nuclear receptor type 1 from F. gigantica, which we named FgNR1, and we hypothesized that the FgNR1 mediates the parasite's response to bile salts. However, the specific ligands are uncharacterized. Crucially, the presence of the signature sequence (Tτ) in the ligand-binding domain (LBD) of FgNR1 suggests that it functions as a transcription factor which will be activated by a specific ligand. This study aims to identify specific ligands for FgNR1 and processes to further identify of antagonistic ligands. The cDNA encoding FgNR1 ligand binding domain (FgNR1-LBD) was cloned, and the recombinant FgNR1-LBD (rFgNR1-LBD) was produced in a prokaryotic expression system. rFgNR1-LBD was used to produced specific polyclonal antibodies for detecting the native protein in the parasite antigen. A multiple sequence alignment of FgNR1-LBD revealed high homology with bile salt responsive nuclear receptors in mammals, suggesting the presence of conserved structural features necessary for ligand recognition and receptor function. Moreover, a luciferase reporter assay demonstrated that bile salts activated FgNR1-LBD functions. Furthermore, molecular docking analysis indicated a strong binding affinity between bile salts and the FgNR1-LBD binding pocket. This study provides important evidence that FgNR1-LBD is functionally responsive to bile salts. Our findings suggest that a critical insight into a potential signaling pathway is pivotal for F. gigantica survival and development within the bile duct. Therefore, FgNR1-LBD represents a novel and promising drug target for developing targeted anti-parasite therapies to block this signaling pathway and improve disease management. |
