2023-12-18 マサチューセッツ大学アマースト校
◆従来の抗生物質は効果を発揮するために水に溶けやすく、かつ油性である必要があり、新しい抗生物質を見つける難しさがあります。病原体が抗生物質に抵抗する仕組みを迎え撃つ新しい戦略として、Type 3分泌システムの機構を妨害することで、病原体を効果的に無力化し、公衆衛生の向上に寄与する可能性があります。
<関連情報>
- https://www.umass.edu/news/article/team-led-umass-amherst-discovers-how-sabotage-antibiotic-resistant-superbugs
- https://pubs.acs.org/doi/10.1021/acsinfecdis.3c00482
スプリットルシフェラーゼを用いた緑膿菌の3型分泌系トランスロコンのアセンブリーを決定する細胞ベースのアッセイ法 Cell-Based Assay to Determine Type 3 Secretion System Translocon Assembly in Pseudomonas aeruginosa Using Split Luciferase
Hanling Guo, Emily J. Geddes, Timothy J. Opperman, and Alejandro P. Heuck
ACS Infectious Diseases Published:November 18, 2023
DOI:https://doi.org/10.1021/acsinfecdis.3c00482
Abstract
Multi-drug-resistant Pseudomonas aeruginosa poses a serious threat to hospitalized patients. This organism expresses an arsenal of virulence factors that enables it to readily establish infections and disseminate in the host. The Type 3 secretion system (T3SS) and its associated effectors play a crucial role in the pathogenesis of P. aeruginosa, making them attractive targets for the development of novel therapeutic agents. The T3SS translocon, composed of PopD and PopB, is an essential component of the T3SS secretion apparatus. In the properly assembled translocon, the N-terminus of PopD protrudes into the cytoplasm of the target mammalian cell, which can be exploited as a molecular indicator of functional translocon assembly. In this article, we describe a novel whole-cell-based assay that employs the split NanoLuc luciferase detection system to provide a readout for translocon assembly. The assay demonstrates a favorable signal/noise ratio (13.6) and robustness (Z′ = 0.67), making it highly suitable for high-throughput screening of small-molecule inhibitors targeting T3SS translocon assembly.
