4TY9

An Ligand-observed Mass Spectrometry-based Approach Integrated into the Fragment Based Lead Discovery Pipeline

4TY9 の概要

• エントリーDOI: 10.2210/pdb4ty9/pdb
• 関連するPDBエントリー: 4TXF 4TXS 4TY8 4TYA 4TYB
• 分子名称: Polyprotein, 5-(trifluoromethyl)pyridin-2-amine (2 entities in total)
• 機能のキーワード: ns5b, tranaferase-transferase inhibitor complex, tranaferase/transferase inhibitor
• 由来する生物種: Hepatitis C virus
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 251230.08

構造登録者

Shui, W.,Yang, C.,Lin, J.,Chen, X.,Qin, S.,Chen, S. (登録日: 2014-07-08, 公開日: 2015-05-06, 最終更新日: 2024-03-20)

主引用文献

Chen, X.,Qin, S.,Chen, S.,Li, J.,Li, L.,Wang, Z.,Wang, Q.,Lin, J.,Yang, C.,Shui, W.
A ligand-observed mass spectrometry approach integrated into the fragment based lead discovery pipeline
Sci Rep, 5:8361-8361, 2015

PubMed Abstract: In fragment-based lead discovery (FBLD), a cascade combining multiple orthogonal technologies is required for reliable detection and characterization of fragment binding to the target. Given the limitations of the mainstream screening techniques, we presented a ligand-observed mass spectrometry approach to expand the toolkits and increase the flexibility of building a FBLD pipeline especially for tough targets. In this study, this approach was integrated into a FBLD program targeting the HCV RNA polymerase NS5B. Our ligand-observed mass spectrometry analysis resulted in the discovery of 10 hits from a 384-member fragment library through two independent screens of complex cocktails and a follow-up validation assay. Moreover, this MS-based approach enabled quantitative measurement of weak binding affinities of fragments which was in general consistent with SPR analysis. Five out of the ten hits were then successfully translated to X-ray structures of fragment-bound complexes to lay a foundation for structure-based inhibitor design. With distinctive strengths in terms of high capacity and speed, minimal method development, easy sample preparation, low material consumption and quantitative capability, this MS-based assay is anticipated to be a valuable addition to the repertoire of current fragment screening techniques.

PubMed: 25666181
DOI: 10.1038/srep08361

実験手法

X-RAY DIFFRACTION (2.78 Å)