Binding hotspots of BAZ2B bromodomain: Histone interaction revealed by solution NMR driven docking.

Fleur M. Ferguson,David M. Dias,Chris Abell,Alessio Ciulli,Hans Wienk,Alexandre M. J. J. Bonvin,Rolf Boelens,João P. G. L. M. Rodrigues

doi:10.1021/bi500909d

Fleur M. Ferguson, David M. Dias + Show 6 more

Open Access

https://doi.org/10.1021/bi500909d

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Abstract

Bromodomains are epigenetic reader domains, which have come under increasing scrutiny both from academic and pharmaceutical research groups. Effective targeting of the BAZ2B bromodomain by small molecule inhibitors has been recently reported, but no structural information is yet available on the interaction with its natural binding partner, acetylated histone H3K14ac. We have assigned the BAZ2B bromodomain and studied its interaction with H3K14ac acetylated peptides by NMR spectroscopy using both chemical shift perturbation (CSP) data and clean chemical exchange (CLEANEX-PM) NMR experiments. The latter was used to characterize water molecules known to play an important role in mediating interactions. Besides the anticipated Kac binding site, we consistently found the bromodomain BC loop as hotspots for the interaction. This information was used to create a data-driven model for the complex using HADDOCK. Our findings provide both structure and dynamics characterization that will be useful in the quest for potent and selective inhibitors to probe the function of the BAZ2B bromodomain.

Highlights

Bromodomains are epigenetic reader domains, which have come under increasing scrutiny both from academic and pharmaceutical research groups
We consistently found the bromodomain BC loop as hotspots for the interaction. This information was used to create a data-driven model for the complex using HADDOCK. Our findings provide both structure and dynamics characterization that will be useful in the quest for potent and selective inhibitors to probe the function of the BAZ2B bromodomain
Previous studies by Philpott et al based on histone peptide arrays and isothermal titration calorimetry (ITC) characterized histone H3K14ac as the binding partner of the BAZ2B bromodomain.[8]

Summary

Introduction

Bromodomains are epigenetic reader domains, which have come under increasing scrutiny both from academic and pharmaceutical research groups. In order to provide a practical structural tool for mapping ligands in its binding site, we assigned the backbone amide protons of the BAZ2B bromodomain by NMR spectroscopy.

Results

Conclusion