Application of Native ESI-MS to Characterize Interactions between Compounds Derived from Fragment-Based Discovery Campaigns and Two Pharmaceutically Relevant Proteins

SLAS Discovery
Agni F M GavriilidouRenato Zenobi

Abstract

Native electrospray ionization mass spectrometry (ESI-MS) was applied to analyze the binding of compounds generated during fragment-based drug discovery (FBDD) campaigns against two functionally distinct proteins, the X-linked inhibitor of apoptosis protein (XIAP) and cyclin-dependent kinase 2 (CDK2). Compounds of different molecular weights and a wide range of binding affinities obtained from the hits to leads and lead optimization stages of FBDD campaigns were studied, and their dissociation constants (Kd) were measured by native ESI-MS. We demonstrate that native ESI-MS has the potential to be applied to the stages of an FBDD campaign downstream of primary screening for the detection and quantification of protein-ligand binding. Native ESI-MS was used to derive Kd values for compounds binding to XIAP, and the dissociation of the complex between XIAP and a peptide derived from the second mitochondria-derived activator of caspases (SMAC) protein induced by one of the test compounds was also investigated. Affinities of compounds binding to CDK2 gave Kd values in the low nanomolar to low millimolar range, and Kd values generated by MS and isothermal titration calorimetry (ITC) followed the same trend for both proteins. Practical...Continue Reading

References

Dec 17, 2003·Analytical Chemistry·Catherine A KeetchCarol V Robinson
Apr 3, 2004·Nature Reviews. Drug Discovery·Michelle R Arkin, James A Wells
Jul 4, 2006·Nature Reviews. Drug Discovery·Steven A Hofstadler, Kristin A Sannes-Lowery
Apr 5, 2007·Nature Protocols·Helena Hernández, Carol V Robinson
Jun 25, 2009·Mass Spectrometry Reviews·Paul Kebarle, Udo H Verkerk
Aug 31, 2010·Magnetic Resonance in Chemistry : MRC·Roger D FarrantRichard J Upton
Mar 1, 2011·Analytical Biochemistry·Minh-Dao Duong-ThiSten Ohlson
Jun 1, 2009·Nature Chemistry·Christopher W Murray, David C Rees
Jan 1, 2010·Future Medicinal Chemistry·Valerie Vivat HannahSarah Sanglier-Cianférani
Apr 8, 2011·Journal of the American Society for Mass Spectrometry·Lan LiuJohn S Klassen
Sep 29, 2011·Journal of the American Society for Mass Spectrometry·Harry J SterlingEvan R Williams
Dec 14, 2011·Journal of Medicinal Chemistry·Hannah J MapleJohn Crosby
Jun 16, 2012·Biochemistry·Duncan E ScottChris Abell
Jan 26, 2013·Analytical Chemistry·Dragana Cubrilovic, Renato Zenobi
Mar 9, 2013·Nature Protocols·Arthur LaganowskyCarol V Robinson
Oct 15, 2013·Nature Methods·Jonathan T S HopperCarol V Robinson
Aug 20, 2014·Journal of the American Society for Mass Spectrometry·Elena N KitovaJohn S Klassen
Jun 9, 2017·Journal of the American Society for Mass Spectrometry·Katharina RootRenato Zenobi

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Citations

Nov 14, 2019·Rapid Communications in Mass Spectrometry : RCM·Ya-Wen LiXian Wang
Aug 21, 2021·Chemical Reviews·Sem TamaraAlbert J R Heck

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Methods Mentioned

BETA
nuclear
surface
thermal shift
NMR
isothermal titration calorimetry
X-ray
gel filtration

Software Mentioned

qNMR
Origin
PRESAT
MATLAB
MNova
WGATE

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