Reference - Detail
| RRC ID | 56603 |
|---|---|
| Author | Omachi K, Kamura M, Teramoto K, Kojima H, Yokota T, Kaseda S, Kuwazuru J, Fukuda R, Koyama K, Matsuyama S, Motomura K, Shuto T, Suico MA, Kai H. |
| Title | A Split-Luciferase-Based Trimer Formation Assay as a High-throughput Screening Platform for Therapeutics in Alport Syndrome. |
| Journal | Cell Chem Biol |
| Abstract |
Alport syndrome is a hereditary glomerular disease caused by mutation in type IV collagen α3-α5 chains (α3-α5(IV)), which disrupts trimerization, leading to glomerular basement membrane degeneration. Correcting the trimerization of α3/α4/α5 chain is a feasible therapeutic approach, but is hindered by lack of information on the regulation of intracellular α(IV) chain and the absence of high-throughput screening (HTS) platforms to assess α345(IV) trimer formation. Here, we developed sets of split NanoLuc-fusion α345(IV) proteins to monitor α345(IV) trimerization of wild-type and clinically associated mutant α5(IV). The α345(IV) trimer assay, which satisfied the acceptance criteria for HTS, enabled the characterization of intracellular- and secretion-dependent defects of mutant α5(IV). Small interfering RNA-based and chemical screening targeting the ER identified several chemical chaperones that have potential to promote α345(IV) trimer formation. This split luciferase-based trimer formation assay is a functional HTS platform that realizes the feasibility of targeting α345(IV) trimers to treat Alport syndrome. |
| Volume | 25(5) |
| Pages | 634-643.e4 |
| Published | 2018-5-17 |
| DOI | 10.1016/j.chembiol.2018.02.003 |
| PII | S2451-9456(18)30043-6 |
| PMID | 29526710 |
| MeSH | Autoantigens / chemistry* Autoantigens / genetics Collagen Type IV / chemistry* Collagen Type IV / genetics Drug Evaluation, Preclinical / methods* HEK293 Cells High-Throughput Screening Assays / methods Humans Nephritis, Hereditary / drug therapy* Nephritis, Hereditary / genetics Point Mutation Protein Multimerization / drug effects* |
| IF | 6.762 |
| Times Cited | 7 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | News |
| Total number of mentions | 21 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | 293T(RCB2202) HeLa(RCB0007) |