8h63

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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(H)Hs) that bind to InlA with picomolar affinities from an alpaca immune library using the phage display method. These InlA-specific V(H)Hs 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(H)Hs inhibited the invasion of L. monocytogenes into host cells in culture. High-resolution X-ray structure analyses of the complexes of V(H)Hs with InlA revealed that the V(H)Hs bind to the same binding site as E-cadherin against InlA. We conclude that these V(H)Hs have the potential for use as drugs to treat listeriosis.
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(H)Hs) that bind to InlA with picomolar affinities from an alpaca immune library using the phage display method. These InlA-specific V(H)Hs 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(H)Hs inhibited the invasion of L. monocytogenes into host cells in culture. High-resolution X-ray structure analyses of the complexes of V(H)Hs with InlA revealed that the V(H)Hs bind to the same binding site as E-cadherin against InlA. We conclude that these V(H)Hs have the potential for use as drugs to treat listeriosis.
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Anti-InlA single-domain antibodies that inhibit the cell invasion of Listeria monocytogenes.,Yamazaki T, Nagatoishi S, Yamawaki T, Nozawa T, Matsunaga R, Nakakido M, Caaveiro JMM, Nakagawa I, Tsumoto K J Biol Chem. 2023 Sep 14:105254. doi: 10.1016/j.jbc.2023.105254. PMID:37716701<ref>PMID:37716701</ref>
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Anti-InlA single-domain antibodies that inhibit the cell invasion of Listeria monocytogenes.,Yamazaki T, Nagatoishi S, Yamawaki T, Nozawa T, Matsunaga R, Nakakido M, Caaveiro JMM, Nakagawa I, Tsumoto K J Biol Chem. 2023 Oct;299(10):105254. doi: 10.1016/j.jbc.2023.105254. Epub 2023 , Sep 14. PMID:37716701<ref>PMID:37716701</ref>
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>

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Crystal structure of Internalin A from Listeria monocytogenes with nanobody VHH10 bound

PDB ID 8h63

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