Fig. 3: Development of Click-IGC for identification of receptors of ligand mixture.
From: Deciphering intercellular signaling complexes by interaction-guided chemical proteomics
a The secretome is first coupled to the probe 3 and then incubated with Ac4ManNAz labeled cells. After adding Cu(I) as catalyst of CuAAC click reaction for 15 min, cells were subjected to AP–MS analysis. b Metabolic labeling of cell surface proteins using Ac4ManNAz, Ac4GalNAz and Ac4GlcNAz. Data are presented as mean ± SD (n = 3 biological replicates). c Flow cytometric analysis of Ac4ManNAz labeled and unlabeled K562 cells incubated with probe 3 in the absence or presence of different concentration of Cu(I), and then conjugated with Streptavidin-Cy3. d Identification of MET by Click-IGC using HGF-BSA mixture (HGF:BSA = 1:250) as ligand on 1, 0.4, and 0.2 million HeLa cells, respectively. Data are presented as mean ± SD (n = 3 biological replicates). e Click-IGC with HGF-BSA solution (HGF:BSA = 1:1000) as ligand on 0.1 million HeLa cells. The MS/MS count of MET is shown in (d). f Photo-IGC (probe 1 or probe 2) or Click-IGC (probe 3) with HGF-BSA (30 ng:15 μg) or LIF-HSA (30 ng:15 μg) on 1 million HeLa cells. Data are presented as mean ± SD (n = 3 biological replicates). Source data are provided as a Source Data file.
