Molecular Glues and Targeted Protein Degradation

The Y2HChem relies on the reconstitution of a functional transcription factor (TF) followed by the expression of a reporter gene in genetically modified yeast cells. Upon a small molecule-induced PPI (mono or bivalent molecule), the DNA Binding Domain (DBD) of the TF is brought in close proximity to its Activation Domain (AD). This activates the transcription of the HIS3 reporter gene, allowing yeast cells to grow on selective medium lacking histidine.

The technique involves 3 components:

• Your protein of interest fused to the DBD
• Target proteins (fragment) fused to the transcriptional AD (the library)
• A small molecule

We offer a large collection of cDNA and genomic libraries (135+) to accommodate your screening project. The libaries are constructed from tissues, cell lines, primary cells or whole organisms derived from various species including human, rodents, plants, bacteria, etc. Using a patent cell-to-cell mating assay, libraries are screened to saturation, averaging an 8-fold in library coverage. Go to Our Libaries

We can construct new libraries on demand to meet your needs.

For Y2HChem projects dedicated to Protein Degradation with Molecular Glues or bifunctional molecules, we can construct focused E3 ligase libraries (full-length or domain-enriched).

Key-benefits of our technology

• Identification of direct interactions
• Enhanced drug intake thanks to proprietary permeable yeast strain
• Exhaustive and unbiased in vivo screening of highly complex domain libraries
• Putative partners are screened individually in a different yeast colony
• No competition with abundant or strong interacting partners
• High sensitivity transcriptional readout and no washing steps
• Integrated bioinformatics: Includes assigning confidence scores & mapping of the interacting domains
• Flagged false positives
• Flexible screening platform

The results of each Y2HChem screen are summarized in three complementary user-friendly deliverables:

• A Results summary with the identified protein partners, the detailed fragments and our confidence score- Predicted Biological Score, PBS®.
• An Excel® table with the sequences and the Genbank accession number and relevant IDs from e.g. Flybase, Wormbase, TAIR.
• The ‘DomSight’ contains a graphical comparison of the experimental interacting domain for each partner and the Smalles Interacting Domain SID®, with known functional and structural domains

Our scientific project leaders support the interpretation of your results and help you to focus on the most relevant interactions for the best outcome of your project.

After results delivery, we can perform different validation experiments using the same technology:

• 1-by-1 in cell validation assays with selected protein targets identified in the Y2HChem screen to confirm binding induced by the small molecule and check specificity.
• Test side-by-side interactions with active and inactive compounds
• Interaction Domain Mapping (IDM) experiments can be performed to further delineate the minimal domain of interaction.

We have developed complementary high-throughput assays to either stabilize or inhibit PPIs with small molecules.

• Y2HChem cell-based assays in 96- or 384-well plates format using HIS3, LacZ or luciferase reporters (1-to-1 format or focused library of protein preys).
• in vitro HTRF® assays in 96- or 384-well plates format for a given protein-protein interaction.
• Chemical libraries available: FDA approved compounds and PPIChem
• Expertise in ubiquitin and deubiquitin enzymes.