4TY8

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

Summary for 4TY8

• Entry DOI: 10.2210/pdb4ty8/pdb
• Related: 4TX3 4TXF 4TY9 4TYA 4TYB
• Descriptor: Polyprotein, 6-methyl-2H-chromen-2-one (3 entities in total)
• Functional Keywords: ns5b, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• Biological source: Hepatitis C virus; More
• Total number of polymer chains: 4
• Total formula weight: 251433.26

Authors

Shui, W.,Yang, C.,Lin, J.,Chen, X.,Qin, S.,Chen, S. (deposition date: 2014-07-08, release date: 2015-05-06, Last modification date: 2024-03-20)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (2.78 Å)