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Chemoenzymatic Labeling to Visualize Intercellular Contacts Using Lipidated SortaseA.

Satoshi Yamaguchi¹, Ryosuke Ikeda¹, Yuki Umeda¹

¹Department of Chemistry and Biotechnology, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.

Chembiochem : a European Journal of Chemical Biology

|August 17, 2022

Summary

This summary is machine-generated.

This study introduces a novel chemoenzymatic method to label contacting cells for analyzing cell-cell communication. The technique selectively visualizes intercellularly contacting cells in mixtures using fluorescent labeling.

Keywords:

cell recognition cell-cell communication enzymes fluorescent labeling protein lipidation

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Area of Science:

Cell Biology
Biochemistry
Medical Diagnostics

Background:

Analyzing intercellular communication is crucial for cell biology and medicine.
Existing methods for labeling contacting cells are limited.
Developing versatile labeling techniques is essential for understanding cell interactions.

Purpose of the Study:

To develop a simple and versatile chemoenzymatic method for labeling intercellularly contacting cells.
To enable selective visualization of cell-cell contacts in mixed cell populations.
To provide a tool for analyzing intercellular communication.

Main Methods:

Utilized a poly(ethylene glycol)-lipid conjugate for cell-surface anchoring.
Decorated cell surfaces with sortase A (SrtA) and a triglycine peptide.
Employed enzymatic labeling with a fluorescent reagent upon cell contact.

Main Results:

Achieved selective fluorescent labeling of triglycine-modified cells upon contact with SrtA-modified cells.
Confirmed selective labeling using confocal microscopy.
Demonstrated the versatility of the chemoenzymatic labeling approach.

Conclusions:

The developed method offers a promising tool for the selective visualization of intercellularly contacting cells.
This technique facilitates the analysis of cell-cell communication in complex cell mixtures.
The chemoenzymatic labeling approach is adaptable for various cell biological and medical applications.