1. Deubiquitinases as Drug Targets in Neglected Tropical Diseases
Global Challenges Research Fund
Jeremy Mottram York Biology
Tony Wilkinson Chemistry
Ana Paula Lima Federal University of Rio de Janeiro, Brazil
Julio Martin Glaxo-Smith-Kline, Spain
Boris Rodenko UbiQ, Netherlands
Drugs for Negelected Diseases Initiative (DNDi) Switzerland

2. NMTs from Protozoan Parasites and their Human Host
H2N
Protein substrate
Gly N H +
CoA-SH
Multiple substrates
Leishmania
Human
Brannigan et al., J. Mol. Biol. 396, 985 (2010); Brannigan et al., IUCr J. 1, 250 (2014)
Frearson et al., Nature 464, 728 (2010)
Hutton et al., J. Med. Chem. 57, 8664 (2014)
Thinon et al., Nat Commun 5, 4919 (2014)

3. Ubiquitin and Ubiquitination
Isopeptide bond between Lysine on Substrate Protein and C-terminal Glycine of Ub
Subsequent isopeptide linkages between Lysines on proximal Ub and C-term of distal Ub
Komander & Rape The Ubiquitin Code Annu Rev Biochem 81, 203 (2012)

4. Polyubiquitin Lys63 tetra-Ub Lys48 tetra-Ub
3HM3; Datta et al., J. Mol. Biol. 392, 1117 (2009)
Lys48 tetra-Ub
2O6V; Eddins et al., J. Mol. Biol. 367, 204 (2007)

5. Ubiquitin Ligases, Ubiquitin Binding Proteins and Deubiquitinases
Substrate
Substrate Ub
Deubiquitinase (DUB) Ub Ub
Downstream Effects Mediated by proteins with Ubiquitin Binding Domains
Protein Degradation – poly Ub(63)
Regulation of Gene Expression – poly Ub(48)

6. Parasite deubiquitinases
DUB – diUb Complex
JC Mottram Laboratory
19 DUBs discovered in Leishmania using activity-based probes
LmDUB1 shown to be essential – also essential in Trypanosoma brucei
1.7 million compounds screened by GSK against TbDUB1 using rhodamine-labelled Ub based fluorescence assay
Qualified hit-list of 14 compounds
Proximal Ub
Distal Ub
5LRV; Mevissen et al., Nature (2016)

7. Plan
Further genetic and chemical validation of DUBs as drug targets
Crystallise and determine crystal structure of LmDUB1 as platform for inhibitor development
Improve chemical properties of inhibitors – potency, selectivity, DMPK
Funding
Outline Application made for MRC GCRF Foundation Award – unsuccessful
Bid for Pump-Priming to UoY BBSRC GCRF – successful – Elena Blagova 4 months 50 % + Consumables + Affinity Reagent