HATRIC-LRC for small molecules
Now available at Dualsystems
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The group of Professor Bernd Wollscheid from ETH Zurich showed in their latest publication that HATRIC-LRC (HATRIC Ligand Receptor Capture) enables the identification of the targets of a small molecule at the cell membrane of living cells to elucidate the mode of action of the molecule. Further, it was demonstrated that the HATRIC-LRC technology platform needs less cells for target identification than the previously published LRC-TriCEPS method and it enables the identification of N-, C-, and O- glycosylated targets.
Target identification using HATRIC-LRC is based on the trifunctional cross linker HATRIC that contains an NHS ester to couple the small molecule, a hydrazone to covalently bind to the carbohydrates of the unknown target proteins at the cell membrane and an azide as pull down function.
The small molecule is coupled to HATRIC using a primary amine. Either your small molecule contains already a primary amine or a primary amine is added by derivatization. In the next step cells expressing the unknown targets and off-targets are mildly oxidized and the HATRIC coupled small molecule is added. The small molecule binds its target(s) at the cell membrane and the second arm of HATRIC covalently binds to the carbohydrates of the target protein(s). During that interaction the cells are still alive. Then the cells are lysed, processed and the third arm of HATRIC is used to pull down the target proteins. Proteins are identified through bottom-up proteomics, relatively quantified and compared to one or several controls. The target candidates are those proteins that are enriched in the ligand of interest samples compared to the control treated samples since the ligand target interaction increases the likelihood of the target carbohydrates to covalently bind to HATRIC compared to the average cell surface protein. Controls can be a positive control such as transferrin or a negative control for example samples treated with an excess of the small molecule devoid of HATRIC prior to adding the HATRIC small molecule conjugate (competition).
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Testimonials from our customers who have used the LRC-TriCEPS technology – in collaboration with Dualsystems Biotech AG.
LRC-TriCEPS customers worldwide
Over 200 satisfied customers from 28 countries.
LRC-TriCEPS publications worldwide
LRC-TriCEPS / HATRIC-LRC Publications
Concerning the LRC-TriCEPS or HATRIC-LRC platforms.
Acidity changes immunology: a new VISTA pathway
Nature Article VISTA interaction with PSGL-1 identified by LRC-TriCEPS
Anti-VISTA antibody that inhibits Vista function and blocks interaction with PSGL-1 and VSIG3 proteins slows tumor growth
Phage resistance at the cost of virulence
PLOS Pathogens | https://doi.org/10.1371/journal.ppat.1008032 October 7, 2019
White Paper Ligand-Receptor Identification Methodologies Details Matter
Validation of extracellular ligand–receptor interactions by Flow‑TriCEPS
Cardiac Targeting Peptide, a Novel Cardiac Vector: Studies in Bio-Distribution, Imaging Application, and Mechanism of Transduction
Leukocyte differentiation by histidine-rich glycoprotein/stanniocalcin-2 complex regulates murine glioma growth through modulation of anti-tumor immunity
Glycomics and Proteomics Approaches to Investigate Early Adenovirus–Host Cell Interactions
HATRIC-based identification of receptors for orphan ligands
Staphylococcal Superantigens Use LAMA2 as a Coreceptor GPCT signaling To Activate T Cells
Toll like receptors TLR1/2, TLR6 and MUC5B as binding interaction partners with cytostatic proline rich polypeptide 1 in human chondrosarcoma
Phenotypic screening—the fast track to novel antibody discovery
Identification of Putative Receptors for the Novel Adipokine CTRP3 Using Ligand-Receptor Capture Technology
Serum stimulation of CCR7 chemotaxis due to coagulation factor XIIa-dependent production of high-molecular-weight kininogen domain 5
Laminin targeting of a peripheral nerve-highlighting peptide enables degenerated nerve visualization
Identification of cell surface receptors for the novel adipokine CTRP3
Dilp8 requires the neuronal relaxin receptor Lgr3 to couple growth to developmental timing
- Received:
- Accepted:
- Published online:
A Mass Spectrometric-Derived Cell Surface Protein Atlas
Protter
Ligand-based receptor identification on living cells and tissues using TRICEPS
Flex your TRICEPS