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Single-domain antibodies as crystallization chaperones to enable structure-based inhibitor development for RBR E3 ubiquitin ligases

DOI: 10.1016/j.chembiol.2019.11.007 DOI Help

Authors: Yi-Chun Isabella Tsai (The Francis Crick Institute) , Henrik Johansson (The Francis Crick Institute; GlaxoSmithKline) , David Dixon (GlaxoSmithKline) , Stephen Martin (The Francis Crick Institute) , Chun-Wa Chung (GlaxoSmithKline) , Jane Clarkson (GlaxoSmithKline) , David House (GlaxoSmithKline) , Katrin Rittinger (The Francis Crick Institute)
Co-authored by industrial partner: Yes

Type: Journal Paper
Journal: Cell Chemical Biology

State: Published (Approved)
Published: December 2019
Diamond Proposal Number(s): 18566

Open Access Open Access

Abstract: Protein ubiquitination plays a key role in the regulation of cellular processes, and misregulation of the ubiquitin system is linked to many diseases. So far, development of tool compounds that target enzymes of the ubiquitin system has been slow and only a few specific inhibitors are available. Here, we report the selection of single-domain antibodies (single-dAbs) based on a human scaffold that recognize the catalytic domain of HOIP, a subunit of the multi-component E3 LUBAC and member of the RBR family of E3 ligases. Some of these dAbs affect ligase activity and provide mechanistic insight into the ubiquitin transfer mechanism of different E2-conjugating enzymes. Furthermore, we show that the co-crystal structure of a HOIP RBR/dAb complex serves as a robust platform for soaking of ligands that target the active site cysteine of HOIP, thereby providing easy access to structure-based ligand design for this important class of E3 ligases.

Journal Keywords: ubiquitination; E3 ubiquitin ligase; crystallization chaperones; domain antibody; covalent inhibitor; fragment screening

Diamond Keywords: Enzymes

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

Instruments: I03-Macromolecular Crystallography , I04-1-Macromolecular Crystallography (fixed wavelength) , I04-Macromolecular Crystallography

Added On: 11/12/2019 11:21


Discipline Tags:

Biochemistry Technique Development - Life Sciences & Biotech Chemistry Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)