Recombinant expression and tryptophan-assisted analysis of human sweet taste receptor T1R3's extracellular domain in sweetener interaction studies
DC Field | Value | Language |
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dc.contributor.author | Jin, Soo-Bin | - |
dc.contributor.author | Kim, Hyun-A | - |
dc.contributor.author | Shin, Ji-Ae | - |
dc.contributor.author | Jung, Na-Hee | - |
dc.contributor.author | Park, Seo-Young | - |
dc.contributor.author | Hong, Sungguan | - |
dc.contributor.author | Kong, Kwang-Hoon | - |
dc.date.accessioned | 2024-04-19T03:00:33Z | - |
dc.date.available | 2024-04-19T03:00:33Z | - |
dc.date.issued | 2024-04 | - |
dc.identifier.issn | 1082-6068 | - |
dc.identifier.issn | 1532-2297 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/73303 | - |
dc.description.abstract | The human palate can discern multiple tastes; however, it predominantly perceives five fundamental flavors: sweetness, saltiness, sourness, bitterness, and umami. Sweetness is primarily mediated through the sweet taste receptor, a membrane-bound heterodimeric structure comprising T1R2-T1R3. However, unraveling the structural and mechanistic intricacies of the sweet taste receptor has proven challenging. This study aimed to address this knowledge gap by expressing an extracellular N-terminal domain encompassing the cysteine-rich domain of human hT1R3 (hT1R3-TMD) in Escherichia coli. The expressed protein was obtained as inclusion bodies, purified by metal affinity chromatography, and refolded using the dilution-refolding method. Through rigorous analysis, we confirmed the successful refolding of hT1R3-TMD and elucidated its structural characteristics using circular dichroism spectroscopy. Notably, the refolded protein was found to exist as either a monomer or a dimer, depending on its concentration. A tryptophan fluorescence quenching assay revealed that the dissociation constants for sucrose, sucralose, and brazzein were >9500 μM, 2380 μM and 14.3 μM, respectively. Our findings highlight the utility of this E. coli expression system for producing functional hT1R3-TMD for investigations and demonstrate the efficacy of the tryptophan fluorescence quenching assay in revealing complex interactions between sweet taste receptors and various sweeteners. | - |
dc.language | 영어 | - |
dc.language.iso | ENG | - |
dc.publisher | TAYLOR & FRANCIS INC | - |
dc.title | Recombinant expression and tryptophan-assisted analysis of human sweet taste receptor T1R3's extracellular domain in sweetener interaction studies | - |
dc.type | Article | - |
dc.identifier.doi | 10.1080/10826068.2024.2336985 | - |
dc.identifier.bibliographicCitation | Preparative biochemistry & biotechnology | - |
dc.description.isOpenAccess | N | - |
dc.identifier.wosid | 001197722300001 | - |
dc.identifier.scopusid | 2-s2.0-85189987474 | - |
dc.citation.title | Preparative biochemistry & biotechnology | - |
dc.type.docType | Article; Early Access | - |
dc.publisher.location | 미국 | - |
dc.subject.keywordAuthor | Expression in Escherichia coli | - |
dc.subject.keywordAuthor | N-terminal domain of hT1R3 | - |
dc.subject.keywordAuthor | human sweet taste receptor | - |
dc.subject.keywordAuthor | refolding process | - |
dc.subject.keywordAuthor | tryptophan assay | - |
dc.subject.keywordPlus | N-TERMINAL DOMAIN | - |
dc.subject.keywordPlus | PROTEIN | - |
dc.subject.keywordPlus | SUBUNITS | - |
dc.subject.keywordPlus | BRAZZEIN | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalResearchArea | Biotechnology & Applied Microbiology | - |
dc.relation.journalWebOfScienceCategory | Biochemical Research Methods | - |
dc.relation.journalWebOfScienceCategory | Biochemistry & Molecular Biology | - |
dc.relation.journalWebOfScienceCategory | Biotechnology & Applied Microbiology | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
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