| RRC ID |
56603
|
| 著者 |
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.
|
| タイトル |
A Split-Luciferase-Based Trimer Formation Assay as a High-throughput Screening Platform for Therapeutics in Alport Syndrome.
|
| ジャーナル |
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.
|
| 巻・号 |
25(5)
|
| ページ |
634-643.e4
|
| 公開日 |
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
|
| 引用数 |
7
|
| リソース情報 |
| ヒト・動物細胞 |
293T(RCB2202)
HeLa(RCB0007) |
