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Fragment Screening against the EthR-DNA Interaction by Native Mass Spectrometry

DOI: 10.1002/anie.201702888 DOI Help

Authors: Daniel Shiu-hin Chan (University of Cambridge) , Vitor Mendes (University of Cambridge) , Sherine E. Thomas (University of Cambridge) , Brendan N. Mcconnell (University of Cambridge) , Dijana Matak-vinković (University of Cambridge) , Anthony G. Coyne (University of Cambridge) , Tom L. Blundell (University of Cambridge) , Chris Abell (University of Cambridge)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Angewandte Chemie International Edition , VOL 277

State: Published (Approved)
Published: May 2017
Diamond Proposal Number(s): 14043

Abstract: Native nanoelectrospray ionization mass spectrometry is an underutilized technique for fragment screening. In this study, the first demonstration is provided of the use of native mass spectrometry for screening fragments against a protein–DNA interaction. EthR is a transcriptional repressor of EthA expression in Mycobacterium tuberculosis (Mtb) that reduces the efficacy of ethionamide, a second-line antitubercular drug used to combat multidrug-resistant Mtb strains. A small-scale fragment screening campaign was conducted against the EthR–DNA interaction using native mass spectrometry, and the results were compared with those from differential scanning fluorimetry, a commonly used primary screening technique. Hits were validated by surface plasmon resonance and X-ray crystallography. The screening campaign identified two new fragments that disrupt the EthR–DNA interaction in vitro (IC50=460–610 μm) and bind to the hydrophobic channel of the EthR dimer.

Journal Keywords: EthR; fragment screening; native mass spectrometry; protein–DNA interactions; tuberculosis

Subject Areas: Biology and Bio-materials, Chemistry, Technique Development

Instruments: I04-Macromolecular Crystallography