HATRIC-based identification of receptors for orphan ligands

• Published in: Nature communications 2018-04, Vol.9 (1), p.1519-8, Article 1519
• Main Authors: Sobotzki, Nadine, Schafroth, Michael A., Rudnicka, Alina, Koetemann, Anika, Marty, Florian, Goetze, Sandra, Yamauchi, Yohei, Carreira, Erick M., Wollscheid, Bernd
• Format: Article
• Language: English
• Subjects: 13/106; 13/31; 14/63; 38/89; 631/154/555; 631/92/96; 82/16; 82/58; Affinity Labels - chemistry; Antibodies - metabolism; Catalysis; Cell Line, Tumor; Cell Membrane - metabolism; Cell surface; Chemical synthesis; Chromatography, Affinity - methods; Click Chemistry - methods; Growth factors; Humanities and Social Sciences; Humans; Influenza A; Influenza A virus - metabolism; Ligands; multidisciplinary; Nutrients; Physiology; Proteomics - methods; Receptors; Receptors, Cell Surface - chemistry; Receptors, Cell Surface - metabolism; Science; Science (multidisciplinary); Solubility; Staining and Labeling - methods; Target recognition; Viruses; Water - chemistry; Workflow
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-018-03936-z

Cellular responses depend on the interactions of extracellular ligands, such as nutrients, growth factors, or drugs, with specific cell-surface receptors. The sensitivity of these interactions to non-physiological conditions, however, makes them challenging to study using in vitro assays. Here we present HATRIC-based ligand receptor capture (HATRIC-LRC), a chemoproteomic technology that successfully identifies target receptors for orphan ligands on living cells ranging from small molecules to intact viruses. HATRIC-LRC combines a click chemistry-based, protein-centric workflow with a water-soluble catalyst to capture ligand-receptor interactions at physiological pH from as few as 1 million cells. We show HATRIC-LRC utility for general antibody target validation within the native nanoscale organization of the surfaceome, as well as receptor identification for a small molecule ligand. HATRIC-LRC further enables the identification of complex extracellular interactomes, such as the host receptor panel for influenza A virus (IAV), the causative agent of the common flu. Technologies for identifying receptor-ligand pairs on living cells at physiological conditions remain scarce. Here, the authors develop a mass spectrometry-based ligand receptor capture technology that can identify receptors for a diverse range of ligands at physiological pH with as few as a million cells.