Time-lapse monitoring of TLR2 ligand internalization with newly developed fluorescent probes

Yohei Arai; Kouhei Yokoyama; Yuki Kawahara; Qi Feng; Ippei Ohta; Atsushi Shimoyama; Shinsuke Inuki; Koichi Fukase; Kazuya Kabayama; Yukari Fujimoto

doi:10.1039/C7OB03205F

Time-lapse monitoring of TLR2 ligand internalization with newly developed fluorescent probes†

Yohei Arai,^a Kouhei Yokoyama,^b Yuki Kawahara,^b Qi Feng,^b Ippei Ohta,^a Atsushi Shimoyama,

^b Shinsuke Inuki,‡^a Koichi Fukase,

^b Kazuya Kabayama

*^b and Yukari Fujimoto

*^a

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* Corresponding authors

^a Faculty of Science and Technology, Keio University, Hiyoshi 3-14-1, Yokohama, Kanagawa 223-8522, Japan
E-mail: fujimotoy@chem.keio.ac.jp

^b Graduate School of Science, Osaka University, Machikaneyama 1-1, Toyonaka, Osaka 560-0043, Japan
E-mail: kaba@chem.sci.osaka-u.ac.jp

Abstract

As a mammalian toll-like receptor family member protein, TLR2 recognizes lipoproteins from bacteria and modulates the immune response by inducing the expression of various cytokines. We have developed fluorescence-labeled TLR2 ligands with either hydrophilic or hydrophobic fluorescence groups. The labeled ligands maintained the inflammatory IL-6 induction activity and enabled us to observe the internalization and colocalization of the TLR2 ligands using live-cell imaging. The time-lapse monitoring in the live-cell imaging of the fluorescence-labeled TLR2 ligand showed that TLR2/CD14 expression in the host cells enhanced the internalization of TLR2 ligand molecules.

This article is part of the themed collection: Celebrating excellence in research: women of organic chemistry

Organic & Biomolecular Chemistry

Time-lapse monitoring of TLR2 ligand internalization with newly developed fluorescent probes†

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Time-lapse monitoring of TLR2 ligand internalization with newly developed fluorescent probes

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