Anti-InlA single-domain antibodies that inhibit the cell invasion of Listeria monocytogenes

Taichi Yamazaki; Satoru Nagatoishi; Tsukushi Yamawaki; Takashi Nozawa; Ryo Matsunaga; Makoto Nakakido; Jose M M Caaveiro; Ichiro Nakagawa; Kouhei Tsumoto

doi:10.1016/j.jbc.2023.105254

Anti-InlA single-domain antibodies that inhibit the cell invasion of Listeria monocytogenes

J Biol Chem. 2023 Oct;299(10):105254. doi: 10.1016/j.jbc.2023.105254. Epub 2023 Sep 14.

Authors

Taichi Yamazaki¹, Satoru Nagatoishi², Tsukushi Yamawaki³, Takashi Nozawa⁴, Ryo Matsunaga¹, Makoto Nakakido¹, Jose M M Caaveiro⁵, Ichiro Nakagawa⁴, Kouhei Tsumoto⁶

Affiliations

¹ Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo, Japan.
² Medical Device Development and Regulation Research Center, School of Engineering, The University of Tokyo, Tokyo, Japan. Electronic address: s-nagatoishi@g.ecc.u-tokyo.ac.jp.
³ Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Tokyo, Japan.
⁴ Department of Microbiology, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
⁵ Laboratory of Global Healthcare, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
⁶ Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo, Japan; Medical Device Development and Regulation Research Center, School of Engineering, The University of Tokyo, Tokyo, Japan; Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Tokyo, Japan. Electronic address: tsumoto@bioeng.t.u-tokyo.ac.jp.

Abstract

Listeriosis, caused by infection with Listeria monocytogenes, is a severe disease with a high mortality rate. The L. monocytogenes virulence factor, internalin family protein InlA, which binds to the host receptor E-cadherin, is necessary to invade host cells. Here, we isolated two single-domain antibodies (V_HHs) that bind to InlA with picomolar affinities from an alpaca immune library using the phage display method. These InlA-specific V_HHs inhibited the binding of InlA to the extracellular domains of E-cadherin in vitro as shown by biophysical interaction analysis. Furthermore, we determined that the V_HHs inhibited the invasion of L. monocytogenes into host cells in culture. High-resolution X-ray structure analyses of the complexes of V_HHs with InlA revealed that the V_HHs bind to the same binding site as E-cadherin against InlA. We conclude that these V_HHs have the potential for use as drugs to treat listeriosis.

Keywords: InlA; Listeria monocytogenes; isothermal titration calorimetry (ITC); listeriosis; protein-protein interaction; single-domain V(H)H antibody; surface plasmon resonance (SPR).