Use of intercellular proximity labeling to quantify and decipher cell-cell interactions directed by diversified molecular pairs

Abstract

FucoID is an intercellular proximity labeling technique for studying cell-cell interactions (CCIs) via fucosyltransferase (FT)–meditated fucosyl-biotinylation, which has been applied to probe antigen-specific dendritic cell (DC)–T cell interactions. In this system, bait cells of interest with cell surface–anchored FT are used to capture the interacting prey cells by transferring a biotin-modified substrate to prey cells. Here, we leveraged FucoID to study CCIs directed by different molecular pairs, e.g., programmed cell death protein-1(PD-1)/programmed cell death protein-ligand-1 (PD-L1), and identify unknown or little studied CCIs, e.g., the interaction of DCs and B cells. To expand the application of FucoID to complex systems, we also synthesized site-specific antibody-based FT conjugate, which substantially improves the ability of FucoID to probe molecular signatures of specific CCI when cells of interest (bait cells) cannot be purified, e.g., in clinical samples. Collectively, these studies demonstrate the general applicability of FucoID to study unknown CCIs in complex systems at a molecular resolution.

GlycoID Proximity Labeling to Identify O‐GlcNAcylated Protein Interactomes in Live Cells

“…Sialic acids on N‐linked glycans have been successfully labeled by linking sialic acid lectins (Siglecs) with metallaphotoredox MicroMap catalysts in a cell surface labeling technique called GlycoMap (Meyer et al., 2022). Finally, another cell‐surface O‐linked glycan labeling technique involving a fucosyltransferase (FucoID) has been developed to capture cell‐cell interactions mediated by glycoprotein recognition (Qiu et al., 2022). A common theme of cell surface glycoprotein proximity labeling is their on‐cell nature, which eliminates some cell permeability concerns.…”

Decoding the Complexity of Immune–Cancer Cell Interactions: Empowering the Future of Cancer Immunotherapy

“…FucoID has several advantages over other proximity labeling approaches. It enables the labeling of glycoproteins, which are an important class of proteins involved in many biological processes [ 36 , 37 ]. Additionally, the fucose tag is relatively small, and it interferes minimally with the function of the labeled proteins, which can reduce the likelihood of introducing artifacts into downstream analyses.…”

Chemical biology tools for protein labelling: insights into cell–cell communication

“…They applied this approach, termed enzyme-mediated proximity cell labelling (EXCELL, Figure 6c ), to the CD40-CD40L interaction. PUP-IT, which uses a ligase fused to a protein of interest to transfer a small protein Pup-E to neighbouring proteins [ 60 ], and Fuco-ID [ 61 , 62 ], which uses a fucosyltransferase to modify cell surface sugars with fucose-biotin, are other contact-dependent methods that have been applied to cell–cell interactions, again in immune signalling. Proximity labelling has also been applied to examine intracellular signalling cascades, for example to monitor O -GlcNAc modification of proteins in response to insulin [ 63 ].…”