Methods to label intercellular contact have attracted attention because of their potential in cell biological and medical applications for the analysis of intercellular communications. In this study, a simple and versatile method for chemoenzymatic labeling of intercellularly contacting cells is demonstrated using a cell‐surface anchoring reagent of a poly(ethylene glycol)(PEG)‐lipid conjugate. The surface of each cell in the cell pairs of interest were decorated with sortase A (SrtA) and triglycine peptide that were lipidated with PEG‐lipid. In the mixture of the two‐cell populations, the triglycine‐modified cells were enzymatically labeled with a fluorescent labeling reagent when in contact with SrtA‐modified cells on a substrate. The selective labeling of the contacting cells was confirmed by confocal microscopy. The method is a promising tool for selective visualization of intercellularly contacting cells in cell mixtures for cell‐cell communication analysis.
Methods to label intercellular contact attract particular attention due to their potential in cell biological and medical applications through analysis of intercellular communications. In this study, a simple and versatile method for chemoenzymatically labeling the intercellularly contacting cell was developed by using a cell-surface anchoring reagent of poly(ethylene glycol)(PEG)-lipid conjugate. The surfaces of each cell in cell pairs of interest were efficiently decorated with sortase A (SrtA) and triglycine peptide that were lipidated with PEG-lipid, respectively. In the mixture of the two cell populations, the triglycine-modified cells were enzymatically labeled with a fluorescent labeling reagent by contacting with the SrtA-modified cells both on the substrate and in cell suspensions. Such selective labeling of the contacting cells was confirmed by confocal microscopy and flow cytometry. The results show a proof of principle that the present method is a promising tool for selective visualization and quantification of the intercellularly contacting cells among cell mixtures in cell-cell communication analysis.
A simple and versatile strategy is introduced for the selective fluorescent labeling of intercellular contacting cells. A transpeptidase sortase A and its substrate triglycine peptide (GGGYC sequence) were chemically conjugated with a lipidated polymer and then anchored onto the membranes of different cells. The triglycine modified cells (displayed in red) were labeled with a fluorescent dye by contacting with the sortase A‐modified cells based on proximity‐dependent transpeptidation with a dye‐modified labeling peptide (DLPETGG sequence). More information can be found in the Research Article by S. Yamaguchi, A. Okamoto et al.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.