Taichi Yamazaki, Satoru Nagatoishi, Tsukushi Yamawaki, Takashi Nozawa, Ryo Matsunaga, Makoto Nakakido, Jose M M Caaveiro, Ichiro Nakagawa, Kouhei Tsumoto
{"title":"抗InlA单结构域抗体,可抑制单核细胞增多性李斯特菌的细胞侵袭。","authors":"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","DOIUrl":null,"url":null,"abstract":"<p><p>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<sub>H</sub>Hs) that bind to InlA with picomolar affinities from an alpaca immune library using the phage display method. These InlA-specific V<sub>H</sub>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<sub>H</sub>Hs inhibited the invasion of L. monocytogenes into host cells in culture. High-resolution X-ray structure analyses of the complexes of V<sub>H</sub>Hs with InlA revealed that the V<sub>H</sub>Hs bind to the same binding site as E-cadherin against InlA. We conclude that these V<sub>H</sub>Hs have the potential for use as drugs to treat listeriosis.</p>","PeriodicalId":22621,"journal":{"name":"The Journal of Biological Chemistry","volume":" ","pages":"105254"},"PeriodicalIF":0.0000,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/79/38/main.PMC10582769.pdf","citationCount":"0","resultStr":"{\"title\":\"Anti-InlA single-domain antibodies that inhibit the cell invasion of Listeria monocytogenes.\",\"authors\":\"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\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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<sub>H</sub>Hs) that bind to InlA with picomolar affinities from an alpaca immune library using the phage display method. These InlA-specific V<sub>H</sub>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<sub>H</sub>Hs inhibited the invasion of L. monocytogenes into host cells in culture. High-resolution X-ray structure analyses of the complexes of V<sub>H</sub>Hs with InlA revealed that the V<sub>H</sub>Hs bind to the same binding site as E-cadherin against InlA. We conclude that these V<sub>H</sub>Hs have the potential for use as drugs to treat listeriosis.</p>\",\"PeriodicalId\":22621,\"journal\":{\"name\":\"The Journal of Biological Chemistry\",\"volume\":\" \",\"pages\":\"105254\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/79/38/main.PMC10582769.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"The Journal of Biological Chemistry\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1016/j.jbc.2023.105254\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2023/9/14 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"The Journal of Biological Chemistry","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.jbc.2023.105254","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/9/14 0:00:00","PubModel":"Epub","JCR":"","JCRName":"","Score":null,"Total":0}
Anti-InlA single-domain antibodies that inhibit the cell invasion of Listeria monocytogenes.
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 (VHHs) that bind to InlA with picomolar affinities from an alpaca immune library using the phage display method. These InlA-specific VHHs 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 VHHs inhibited the invasion of L. monocytogenes into host cells in culture. High-resolution X-ray structure analyses of the complexes of VHHs with InlA revealed that the VHHs bind to the same binding site as E-cadherin against InlA. We conclude that these VHHs have the potential for use as drugs to treat listeriosis.