Yeast Two-Hybrid basedProtein Interactions screening

Our ULTImate Y2H™ is the most comprehensive and in-depth yeast two-hybrid system identifying protein-protein interactions as well as DNA, RNA and bioactive small molecule interactions.

Our Technology

Our ULTImate yeast two-hybrid (Y2H) screening platform

Our optimized ULTImate Y2H™ technology takes advantage of our patented cell-to-cell mating process which permits the testing of ~ 83 million interactions per screen, ensuring that our high-complexity domain-enriched libraries are screened to saturation and with high reproducibility. As a result, rare binding partners and transient interactions are identified.


Included in our deliverables and integrated bioinformatic tools, we compute confidence scores allowing you to focus on the most relevant interactions. Technical false positives and highly connected proteins are flagged.

Please have a look at our Ressources Center for the basics of the Y2H principle.

Our Libraries

Our ULTImate libraries

We are proud to offer a large collection of over 135 libraries from 45+ species, including human, mouse, insect, plant and bacteria.

Each ULTImate cDNA library is validated using stringent quality-control requirements:

  • Library complexity is at least 10 million independent clones
  • Insert size distribution is between 800-1000 bp, which corresponds to the average size of a protein domain
  • Ribosomal RNA, mitochondrial DNA and empty vector represents less than 10% of the library clones


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

Our solutions

ULTImate Yeast Two-Hybrid

ULTImate Yeast Two-Hybrid Y2H
MBmate Y2H for Membrane Proteins
ULTImate Y1H for DNA
ULTImate Y3H for RNA
ULTImate Y2H + Co-factor(s)
Loss of Affinity Mutants
1 by 1 Interaction Domain Mapping and Validation
Y2H NGS - Next-Generation Sequencing
Deliverables
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Molecules tested: Soluble proteins

  • Full-length proteins, fragments or peptides
  • Cytoplasmic and secreted proteins
  • Loop regions, cytoplasmic tails and extracellular domains of membrane proteins

Key benefits

  • Rapid & exhaustive screening of high-complexity domain-enriched libraries
  • Identification of weak and rare binding partners
  • Up to 380 positive clones sequenced (5' & 3') in a single screen
  • Comprehensive & integrated bioinformatics analysis of results and annotation of functional, structural and interacting domains of the interacting proteins
  • Back-up screening strategies included
  • High-quality scientific & technical assistance regarding bait design and screening strategy for the best possible outcome
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Molecules tested: Membrane-associated proteins

  • Full-length and fragments of transmembrane-containing proteins

Key benefits

  • Identify protein targets of single or multipass membrane proteins
  • Functional assays include bait testing for proper localization & orientation
  • Rapid & exhaustive screening of custom-made MBmate libraries
  • Up to 380 positive clones sequenced
  • High-quality scientific & technical assistance regarding bait design and screening strategy for the best possible outcome 
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Molecules Tested: DNA sequences (i.e., promoter regions, responsive elements)

Key benefits

  • Rapid & exhaustive screening of high-complexity domain-enriched libraries
  • Up to 380 positive clones sequenced (5'& 3') in a single screen
  • Comprehensive & integrated bioinformatics analysis of results and annotation of functional, structural and interacting domains of the interacting proteins
  • Back-up screening strategies included
  • High-quality scientific & technical assistance regarding bait design and screening strategy for the best possible outcome
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Molecules Tested: RNA sequences

Key Benefits

  • Robust MS-2 bait system
  • RNA baits can be screened in both 5’ and 3’ orientations
  • Up to 380 positive clones sequenced (5'& 3') in a single screen
  • Comprehensive & integrated bioinformatics analysis of results and annotation of functional, structural and interacting domains of the interacting proteins
  • Project team leader provides scientific & technical assistance regarding bait design and screening strategy for the best possible outcome
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The ULTImate Y2H+1 screen is a modified version of the ULTImate Y2H system where the expression of a co-factor in yeast is required for either the proper folding (i.e., chaperone) or post-translational modification (i.e., tyrosine kinase) of the bait. Co-expression of several proteins of a complex is also possible.

 

Molecule tested: Soluble protein + co-factor 

Key Benefits

  • Rapid & exhaustive screening of high-complexity domain-enriched libraries
  • Identification of weak and rare binding partners
  • Comprehensive & integrated bioinformatics analysis of results and annotation of functional, structural and interacting domains of the interacting proteins
  • Project team leader provides scientific & technical assistance regarding bait design and screening strategy for the best possible outcome
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Molecules tested: Soluble proteins

  • Full-length proteins, fragments or peptides
  • Cytoplasmic or extracellular proteins
  • Loops and tails of membrane proteins


Key Benefits

  • Robust screening method to identify single point mutation(s) that are responsible for the protein-protein interaction (PPI) based on ULTImate Y2H™
  • In-depth understanding of PPI interface at the single amino acid level
  • Generation of invaluable tools for functional studies
  • Exploit results to conduct an ULTImate Y2H™ screen or 1by1 Y2H assays

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Our 1by1 interaction service features: 1) validating the interaction between two proteins identified in a Y2H screen or by other approaches, 2) mapping the interacting domain of a protein and 3) comparing the binding of wild-type and mutant proteins or between isoforms.


Molecules Tested:

  • Wild-type and mutant full-length proteins
  • Protein fragments and peptides
  • Cytoplasmic and extracellular proteins
  • Loops, tails and membrane-associated proteins

Key-benefits

  • Simple, fast and robust pairwise interaction assays
  • In-depth characterization of the PPI and domains involved
  • Publication-grade illustrations including all positive and negative controls
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We provide our customers with Next Generation Sequencing (NGS) analysis to accommodate their needs.

While traditional Sanger sequencing is the most reliable and qualitative tool for the majority of our Y2H projects there are exceptions.
Some baits naturally interact with several highly abundant targets in a Y2H screen, which can obscure the detection of low affinity and/or rare protein binding partners. The selection pressure is increased to reduce the number of positive clones that can be analyzed by standard sequencing, which can result in the loss of low affinity and rare interactions. In such a case, we propose NGS in order to uncover all potential interactors.

Key Benefits

  • Generation of high-quality NGS dataset; no articifial amplication of background
  • Low affinity and low abundant interactions identified
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Our deliverables are presented and organized in a user-friendly format.

The results of each ULTImate screen contain three complementary files:

A Results summary with the identified protein partners, the detailed fragments and our confidence score- the 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 Smallest Interacting Domain SID®, with known functional and structural domains.

our publications

Publications

The oncogene AAMDC links PI3K-AKT-mTOR signaling with metabolic reprograming in estrogen receptor-positive breast cancer

Golden E, Rashwan R, Woodward EA, Sgro A, Wang E, Sorolla A, Waryah C, Tie WJ, Cuyàs E, Ratajska M, Kardaś I, Kozlowski P, Johnstone EKM, See HB, Duffy C, Parry J, Lagerborg KA, Czapiewski P, Menendez JA, Gorczyński A, Wasag B, Pfleger KDG, Curtis C, Lee BK, Kim J, Cursons J, Pavlos NJ, Biernat W, Jain M, Woo AJ. Redfern A, Blancafort P (2021) Nat Commun, 12(1):1920

Parasitic modulation of host development by ubiquitin-independent protein degradation

Huang W, MacLean AM, Sugio A, Maqbool A, Busscher M, Cho ST, Kamoun S, Kuo CH, Immink RGH, Hogenhout. SA (2021) Cell, 184(20):5201-5214.e12

JASMONATE-ZIM DOMAIN proteins engage Polycomb chromatin modifiers to modulate Jasmonate signaling in Arabidopsis

Li Z, Luo X, Ou Y, Jiao H, Peng L, Fu X, Macho AP, Liu R, He Y Mol Plant (2021) 3;14(5):732-747

FAQ

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