Cargando…

Human LYPD8 protein inhibits motility of flagellated bacteria

BACKGROUND: We previously reported that the mouse Ly6/Plaur domain containing 8 (mLypd8), a GPI-anchored protein highly and selectively expressed on colonic epithelia, contributes to segregation of intestinal microbiota and intestinal epithelia and is critical for prevention of intestinal inflammati...

Descripción completa

Detalles Bibliográficos
Autores principales: Hsu, Chiao-Ching, Okumura, Ryu, Takeda, Kiyoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5725809/
https://www.ncbi.nlm.nih.gov/pubmed/29259722
http://dx.doi.org/10.1186/s41232-017-0056-3
_version_ 1783285607429046272
author Hsu, Chiao-Ching
Okumura, Ryu
Takeda, Kiyoshi
author_facet Hsu, Chiao-Ching
Okumura, Ryu
Takeda, Kiyoshi
author_sort Hsu, Chiao-Ching
collection PubMed
description BACKGROUND: We previously reported that the mouse Ly6/Plaur domain containing 8 (mLypd8), a GPI-anchored protein highly and selectively expressed on colonic epithelia, contributes to segregation of intestinal microbiota and intestinal epithelia and is critical for prevention of intestinal inflammation. In addition, it was found that human LYPD8 (hLYPD8) is expressed in the colonic epithelia and expression of hLYPD8 is reduced in some ulcerative colitis patients. However, the molecular characteristics and functions of hLYPD8 remain unclear. In this study, we generated the hLYPD8 protein and characterized its functions. METHODS: To analyze the characteristics and functions of the hLYPD8 protein, recombinant FLAG-tagged hLYPD8 protein was generated by two kinds of protein expression systems: a mammalian cell expression system and a Pichia pastoris expression system. Recombinant hLYPD8 protein was analyzed by western blot analysis or deglycosylation assay. The effect of the protein on flagellated bacteria was examined by ELISA assay and motility assay using semi-agar plates. RESULTS: hLYPD8 was a highly N-glycosylated GPI-anchored protein, like mLypd8. Moreover, recombinant hLYPD8 protein generated by the Pichia pastoris expression system using the SuperMan(5) strain, which enabled production of a large number of proteins with human-like glycosylation, presented the high binding affinity and the motility inhibitory function to flagellated bacteria, such as Proteus mirabilis. CONCLUSIONS: These results demonstrated that hLYPD8 inhibits the motile activity of flagellated bacteria, many of which are involved in intestinal inflammation. The supplementation of recombinant hLYPD8 protein might be a novel therapeutic approach for intestinal inflammation of inflammatory bowel diseases.
format Online
Article
Text
id pubmed-5725809
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-57258092017-12-19 Human LYPD8 protein inhibits motility of flagellated bacteria Hsu, Chiao-Ching Okumura, Ryu Takeda, Kiyoshi Inflamm Regen Research Article BACKGROUND: We previously reported that the mouse Ly6/Plaur domain containing 8 (mLypd8), a GPI-anchored protein highly and selectively expressed on colonic epithelia, contributes to segregation of intestinal microbiota and intestinal epithelia and is critical for prevention of intestinal inflammation. In addition, it was found that human LYPD8 (hLYPD8) is expressed in the colonic epithelia and expression of hLYPD8 is reduced in some ulcerative colitis patients. However, the molecular characteristics and functions of hLYPD8 remain unclear. In this study, we generated the hLYPD8 protein and characterized its functions. METHODS: To analyze the characteristics and functions of the hLYPD8 protein, recombinant FLAG-tagged hLYPD8 protein was generated by two kinds of protein expression systems: a mammalian cell expression system and a Pichia pastoris expression system. Recombinant hLYPD8 protein was analyzed by western blot analysis or deglycosylation assay. The effect of the protein on flagellated bacteria was examined by ELISA assay and motility assay using semi-agar plates. RESULTS: hLYPD8 was a highly N-glycosylated GPI-anchored protein, like mLypd8. Moreover, recombinant hLYPD8 protein generated by the Pichia pastoris expression system using the SuperMan(5) strain, which enabled production of a large number of proteins with human-like glycosylation, presented the high binding affinity and the motility inhibitory function to flagellated bacteria, such as Proteus mirabilis. CONCLUSIONS: These results demonstrated that hLYPD8 inhibits the motile activity of flagellated bacteria, many of which are involved in intestinal inflammation. The supplementation of recombinant hLYPD8 protein might be a novel therapeutic approach for intestinal inflammation of inflammatory bowel diseases. BioMed Central 2017-12-04 /pmc/articles/PMC5725809/ /pubmed/29259722 http://dx.doi.org/10.1186/s41232-017-0056-3 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Hsu, Chiao-Ching
Okumura, Ryu
Takeda, Kiyoshi
Human LYPD8 protein inhibits motility of flagellated bacteria
title Human LYPD8 protein inhibits motility of flagellated bacteria
title_full Human LYPD8 protein inhibits motility of flagellated bacteria
title_fullStr Human LYPD8 protein inhibits motility of flagellated bacteria
title_full_unstemmed Human LYPD8 protein inhibits motility of flagellated bacteria
title_short Human LYPD8 protein inhibits motility of flagellated bacteria
title_sort human lypd8 protein inhibits motility of flagellated bacteria
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5725809/
https://www.ncbi.nlm.nih.gov/pubmed/29259722
http://dx.doi.org/10.1186/s41232-017-0056-3
work_keys_str_mv AT hsuchiaoching humanlypd8proteininhibitsmotilityofflagellatedbacteria
AT okumuraryu humanlypd8proteininhibitsmotilityofflagellatedbacteria
AT takedakiyoshi humanlypd8proteininhibitsmotilityofflagellatedbacteria