A5014976573- Cancer-related gene regulation
- Peptidase Inhibition and Analysis
- Ubiquitin and proteasome pathways
- RNA modifications and cancer
- Protein Degradation and Inhibitors
- RNA and protein synthesis mechanisms
- DNA and Nucleic Acid Chemistry
- Chemical Synthesis and Analysis
- Epigenetics and DNA Methylation
- Computational Drug Discovery Methods
- Protein Structure and Dynamics
- Glycosylation and Glycoproteins Research
- Enzyme Structure and Function
- Histone Deacetylase Inhibitors Research
- DNA Repair Mechanisms
- Click Chemistry and Applications
- RNA Research and Splicing
- CRISPR and Genetic Engineering
- Amyloidosis: Diagnosis, Treatment, Outcomes
- Cellular transport and secretion
- Synthesis and Characterization of Heterocyclic Compounds
- interferon and immune responses
- Cytokine Signaling Pathways and Interactions
- Receptor Mechanisms and Signaling
- Biochemical and Molecular Research
University of Toronto
2018-2025
Structural Genomics Consortium
2018-2025
Princess Margaret Cancer Centre
2018-2024
University Health Network
2024
Canada Research Chairs
2021
University of Milan
2013-2017
DCAF1 is a substrate receptor of two distinct E3 ligases (CRL4DCAF1 and EDVP), plays critical physiological role in protein degradation, considered drug target for various cancers. Antagonists could be used toward the development therapeutics cancers viral treatments. We WDR domain to screen 114-billion-compound DNA encoded library (DEL) identified candidate compounds using similarity search machine learning. This led discovery compound (Z1391232269) with an SPR KD 11 μM. Structure-guided...
Abstract Eukaryotic elongation factor 1 alpha (eEF1A) delivers aminoacyl-tRNA to the ribosome and thereby plays a key role in protein synthesis. Human eEF1A is subject extensive post-translational methylation, but several of responsible enzymes remain unknown. Using wide range experimental approaches, we here show that human methyltransferase (MTase)-like 13 (METTL13) contains two distinct MTase domains targeting N terminus Lys55 eEF1A, respectively. Our biochemical structural analyses...
Abstract Protein arginine methyltransferases (PRMTs) regulate diverse biological processes and are increasingly being recognized for their potential as drug targets. Here we report the discovery of a potent, selective, cell-active chemical probe PRMT7. SGC3027 is cell permeable prodrug, which in cells converted to SGC8158, SAM-competitive PRMT7 inhibitor. Inhibition or knockout cellular results drastically reduced levels monomethylated HSP70 family stress-associated proteins. Structural...
The CACHE challenges are a series of prospective benchmarking exercises to evaluate progress in the field computational hit-finding. Here we report results inaugural challenge which 23 teams each selected up 100 commercially available compounds that they predicted would bind WDR domain Parkinson's disease target LRRK2, with no known ligand and only an apo structure PDB. lack binding data presumably low druggability is hit finding methods. Of 1955 molecules by participants Round 1 challenge,...
DCAF1 functions as a substrate recruitment subunit for the RING-type CRL4DCAF1 and HECT family EDVPDCAF1 E3 ubiquitin ligases. The WDR domain of serves binding platform proteins is also targeted by HIV SIV lentiviral adaptors to induce ubiquitination proteasomal degradation antiviral host factors. It therefore attractive both potential therapeutic target development chemical inhibitors an ligase that could be recruited novel PROTACs protein degradation. In this study, we used proteome-scale...
Abstract Short-chain acylations of lysine residues in eukaryotic proteins are recognized as essential posttranslational chemical modifications (PTMs) that regulate cellular processes from transcription, cell cycle, metabolism, to signal transduction. Lysine butyrylation was initially discovered a normal straight chain (Knbu). Here we report its structural isomer, branched butyrylation, i.e. isobutyrylation (Kibu), existing new PTM on nuclear histones. Uniquely, isobutyryl-CoA is derived...
Histone lysine crotonylation is a posttranslational modification with demonstrated functions in transcriptional regulation. Here we report the discovery of new type histone modification, methacrylation (Kmea), corresponding to structural isomer crotonyllysine. We validate identity this using diverse chemical approaches and further confirm occurrence mark by pan specific site-specific anti-methacryllysine antibodies. In total, identify 27 Kmea modified sites HeLa cells affinity enrichment...
Abstract Cbl-b is a RING-type E3 ubiquitin ligase that expressed in several immune cell lineages, where it negatively regulates the activity of cells. has specifically been identified as an attractive target for cancer immunotherapy due to its role promoting immunosuppressive tumor environment. A inhibitor, Nx-1607, currently phase I clinical trials advanced solid malignancies. Using suite biophysical and cellular assays, we confirm potent binding C7683 (an analogue Nx-1607) full-length...
ou non, émanant des établissements d'enseignement et de recherche français étrangers, laboratoires publics privés.
Target class-focused drug discovery has a strong track record in pharmaceutical research, yet public domain data indicate that many members of protein families remain unliganded. Here we present systematic approach to scale up the and characterization small molecule ligands for WD40 repeat (WDR) family. We developed comprehensive suite protocols production, crystallography, biophysical, biochemical, cellular assays. A pilot hit-finding campaign using DNA-encoded chemical library selection...
Aberrant isoform expression of chromatin-associated proteins can induce epigenetic programs related to disease. The MDS1 and EVI1 complex locus (MECOM) encodes PRDM3, a protein with an N-terminal PR-SET domain, as well shorter isoform, EVI1, lacking the N-terminus containing domain (ΔPR). Imbalanced MECOM isoforms is observed in multiple malignancies, implicating oncogene, while PRDM3 has been suggested function tumor suppressor through unknown mechanism. To elucidate functional...
PRDM9 is a PR domain containing protein which trimethylates histone 3 on lysine 4 and 36. Its normal expression restricted to germ cells attenuation of its activity results in altered meiotic gene transcription, impairment double-stranded breaks pairing between homologous chromosomes. There growing evidence for role aberrant oncogenesis genome instability. Here we report the discovery MRK-740, potent (IC50: 80 ± 16 nM), selective cell-active inhibitor (Chemical Probe). MRK-740 binds...
Protein arginine methyltransferase 6 (PRMT6) plays important roles in several biological processes associated with multiple cancers. Well-characterized potent, selective, and cell-active PRMT6 inhibitors are invaluable tools for testing therapeutic hypotheses. Although there known reversible inhibitors, covalent have not been reported. Based on a cocrystal structure of PRMT6–MS023 (a type I PRMT inhibitor), we discovered the first potent irreversible inhibitor, 4 (MS117). The binding mode...
Controlling malaria requires new drugs against Plasmodium falciparum. The P. falciparum cGMP-dependent protein kinase (PfPKG) is a validated target whose inhibitors could block multiple steps of the parasite's life cycle. We defined structure–activity relationship (SAR) pyrrole series for PfPKG inhibition. Key pharmacophores were modified to enable full exploration chemical diversity and gain knowledge about an ideal core scaffold. In vitro potency recombinant human PKG used determine...
The human pseudouridine synthase PUS7 is a versatile RNA modification enzyme targeting many RNAs thereby playing critical role in development and brain function. Whereas all target of share consensus sequence, additional recognition elements are likely required, the structural basis for binding by unknown. Here, we characterize structure-function relationship reporting its X-ray crystal structure at 2.26 Å resolution. Compared to bacterial homolog, possesses two subdomains, modeling studies...
WD40 repeat-containing protein 91 (WDR91) regulates early-to-late endosome conversion and plays vital roles in fusion, recycling, transport. WDR91 was recently identified as a potential host factor for viral infection. We employed DNA-encoded chemical library (DEL) selection against the WDR domain of WDR91, followed by machine learning to predict ligands from synthetically accessible Enamine REAL database. Screening predicted compounds selective compound
Critical Assessment of Computational Hit-Finding Experiments (CACHE) Challenges emerged as real-life stress tests for computational hit-finding strategies. In CACHE Challenge #1, 23 participants contributed their original workflows to identify small-molecule ligands the WD40 repeat (WDR) LRRK2, a promising Parkinson's target. We applied FRASE-based robot (FRASE-bot), platform interaction-based screening allowing drastic reduction explorable chemical space and concurrent detection putative...
We report an enantioselective protein affinity selection mass spectrometry screening approach (EAS-MS) that enables the detection of weak binders, informs about selectivity, and generates orthogonal confirmation binding. After method development with control proteins, we screened 31 human proteins against a designed library 8,210 chiral compounds. 16 binders to 12 targets, including many predicted be "challenging ligand", were discovered confirmed in assays. 7 6 targets bound manner, K D s...
Abstract Human DExD/H-box RNA helicases are ubiquitous molecular motors that unwind and rearrange secondary structures in an ATP-dependent manner. These enzymes play essential roles nearly all aspects of metabolism. While their biological functions well-characterized, the kinetic mechanisms remain relatively understudied vitro . In this study, we describe development optimization a bioluminescence-based assay to kinetically characterize three human helicases: MDA5, LGP2, DDX1. The assays...
A promising drug target, SETDB1, is a dual methyl-lysine (Kme) reader and methyltransferase implicated in cancer neurodegenerative disease progression. To help understand the role of triple Tudor domain (3TD) its Kme reader, we first identify low micromolar potency small molecule ligand, UNC6535, which occupies simultaneously both TD2 TD3 binding sites. Further optimization leads to discovery UNC10013, covalent 3TD ligand targeting Cys385 SETDB1. UNC10013 potent with kinact/KI 1.0 × 106...