論文 - 詳細
| RRC ID | 30717 |
|---|---|
| 著者 | Yoshimoto N, Tatematsu K, Iijima M, Niimi T, Maturana AD, Fujii I, Kondo A, Tanizawa K, Kuroda S. |
| タイトル | High-throughput de novo screening of receptor agonists with an automated single-cell analysis and isolation system. |
| ジャーナル | Sci Rep |
| Abstract |
Reconstitution of signaling pathways involving single mammalian transmembrane receptors has not been accomplished in yeast cells. In this study, intact EGF receptor (EGFR) and a cell wall-anchored form of EGF were co-expressed on the yeast cell surface, which led to autophosphorylation of the EGFR in an EGF-dependent autocrine manner. After changing from EGF to a conformationally constrained peptide library, cells were fluorescently labeled with an anti-phospho-EGFR antibody. Each cell was subjected to an automated single-cell analysis and isolation system that analyzed the fluorescent intensity of each cell and automatically retrieved each cell with the highest fluorescence. In ~3.2 × 10(6) peptide library, we isolated six novel peptides with agonistic activity of the EGFR in human squamous carcinoma A431 cells. The combination of yeast cells expressing mammalian receptors, a cell wall-anchored peptide library, and an automated single-cell analysis and isolation system might facilitate a rational approach for de novo drug screening. |
| 巻・号 | 4 |
| ページ | 4242 |
| 公開日 | 2014-2-28 |
| DOI | 10.1038/srep04242 |
| PII | srep04242 |
| 解説 | S.cerevisiaeでGal1プロモーター制御下での目的遺伝子の発現に使用できます。本研究ではアゴニスト探索のためにEGFRを酵母で発現するために使用されました。 |
| PMID | 24577528 |
| PMC | PMC3937795 |
| MeSH | Biological Assay / instrumentation* Biological Assay / methods Cell Separation / instrumentation* Cell Separation / methods Drug Evaluation, Preclinical / instrumentation* Drug Evaluation, Preclinical / methods Equipment Design Equipment Failure Analysis Flow Cytometry / instrumentation* Flow Cytometry / methods High-Throughput Screening Assays / instrumentation* High-Throughput Screening Assays / methods Receptors, Cytoplasmic and Nuclear / antagonists & inhibitors* Robotics / instrumentation* Robotics / methods |
| IF | 3.998 |
| 引用数 | 6 |
| WOS 分野 | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 9 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 遺伝子材料 | pGMH20 (RDB01956). |