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Chemoenzymatic Labeling to Visualize Intercellular Contacts Using Lipidated SortaseA

Von Wiley-VCH zur Verfügung gestellt

A simple and versatile strategy toward selective fluorescent labeling of intercellular contacting cells is reported. A transpeptidase sortase A and its substrate triglycine peptide were conjugated with a lipidated polymer, leading to anchoring onto cell membranes. The triglycine-modified cells were labeled with a fluorescent dye by contacting with the sortase A-modified cells based on proximity-dependent transpeptidation.


Abstract

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.

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