Binding determinants in the interplay between porcine aminopeptidase N and enterotoxigenic Escherichia coli F4 fimbriae
Abstract The binding of F4+ enterotoxigenic Escherichia coli (ETEC) and the specific receptor on porcine intestinal epithelial cells is the initial step in F4+ ETEC infection. Porcine aminopeptidase N (APN) is a newly discovered receptor for F4 fimbriae that binds directly to FaeG adhesin, which is...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2018-02-01
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| Series: | Veterinary Research |
| Online Access: | http://link.springer.com/article/10.1186/s13567-018-0519-9 |
| _version_ | 1818576069370839040 |
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| author | Pengpeng Xia Guomei Quan Yi Yang Jing Zhao Yiting Wang Mingxu Zhou Philip R. Hardwidge Jianzhong Zhu Siguo Liu Guoqiang Zhu |
| author_facet | Pengpeng Xia Guomei Quan Yi Yang Jing Zhao Yiting Wang Mingxu Zhou Philip R. Hardwidge Jianzhong Zhu Siguo Liu Guoqiang Zhu |
| author_sort | Pengpeng Xia |
| collection | DOAJ |
| description | Abstract The binding of F4+ enterotoxigenic Escherichia coli (ETEC) and the specific receptor on porcine intestinal epithelial cells is the initial step in F4+ ETEC infection. Porcine aminopeptidase N (APN) is a newly discovered receptor for F4 fimbriae that binds directly to FaeG adhesin, which is the major subunit of the F4 fimbriae variants F4ab, F4ac, and F4ad. We used overlapping peptide assays to map the APN-FaeG binding sites, which has facilitated in the identifying the APN-binding amino acids that are located in the same region of FaeG variants, thereby limiting the major binding regions of APN to 13 peptides. To determine the core sequence motif, a panel of FaeG peptides with point mutations and FaeG mutants were constructed. Pull-down and binding reactivity assays using piglet intestines determined that the amino acids G159 of F4ab, N209 and L212 of F4ac, and A200 of F4ad were the critical residues for APN binding of FaeG. We further show using ELISA and confocal microscopy assay that amino acids 553–568, and 652–670 of the APN comprise the linear epitope for FaeG binding in all three F4 fimbriae variants. |
| first_indexed | 2024-12-15T00:49:14Z |
| format | Article |
| id | doaj.art-f0a63beb6e5e45feb35d2174ffa05737 |
| institution | Directory Open Access Journal |
| issn | 1297-9716 |
| language | English |
| last_indexed | 2024-12-15T00:49:14Z |
| publishDate | 2018-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Veterinary Research |
| spelling | doaj.art-f0a63beb6e5e45feb35d2174ffa057372022-12-21T22:41:27ZengBMCVeterinary Research1297-97162018-02-0149111110.1186/s13567-018-0519-9Binding determinants in the interplay between porcine aminopeptidase N and enterotoxigenic Escherichia coli F4 fimbriaePengpeng Xia0Guomei Quan1Yi Yang2Jing Zhao3Yiting Wang4Mingxu Zhou5Philip R. Hardwidge6Jianzhong Zhu7Siguo Liu8Guoqiang Zhu9College of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Kansas State UniversityCollege of Veterinary Medicine, Yangzhou UniversityNational Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural SciencesCollege of Veterinary Medicine, Yangzhou UniversityAbstract The binding of F4+ enterotoxigenic Escherichia coli (ETEC) and the specific receptor on porcine intestinal epithelial cells is the initial step in F4+ ETEC infection. Porcine aminopeptidase N (APN) is a newly discovered receptor for F4 fimbriae that binds directly to FaeG adhesin, which is the major subunit of the F4 fimbriae variants F4ab, F4ac, and F4ad. We used overlapping peptide assays to map the APN-FaeG binding sites, which has facilitated in the identifying the APN-binding amino acids that are located in the same region of FaeG variants, thereby limiting the major binding regions of APN to 13 peptides. To determine the core sequence motif, a panel of FaeG peptides with point mutations and FaeG mutants were constructed. Pull-down and binding reactivity assays using piglet intestines determined that the amino acids G159 of F4ab, N209 and L212 of F4ac, and A200 of F4ad were the critical residues for APN binding of FaeG. We further show using ELISA and confocal microscopy assay that amino acids 553–568, and 652–670 of the APN comprise the linear epitope for FaeG binding in all three F4 fimbriae variants.http://link.springer.com/article/10.1186/s13567-018-0519-9 |
| spellingShingle | Pengpeng Xia Guomei Quan Yi Yang Jing Zhao Yiting Wang Mingxu Zhou Philip R. Hardwidge Jianzhong Zhu Siguo Liu Guoqiang Zhu Binding determinants in the interplay between porcine aminopeptidase N and enterotoxigenic Escherichia coli F4 fimbriae Veterinary Research |
| title | Binding determinants in the interplay between porcine aminopeptidase N and enterotoxigenic Escherichia coli F4 fimbriae |
| title_full | Binding determinants in the interplay between porcine aminopeptidase N and enterotoxigenic Escherichia coli F4 fimbriae |
| title_fullStr | Binding determinants in the interplay between porcine aminopeptidase N and enterotoxigenic Escherichia coli F4 fimbriae |
| title_full_unstemmed | Binding determinants in the interplay between porcine aminopeptidase N and enterotoxigenic Escherichia coli F4 fimbriae |
| title_short | Binding determinants in the interplay between porcine aminopeptidase N and enterotoxigenic Escherichia coli F4 fimbriae |
| title_sort | binding determinants in the interplay between porcine aminopeptidase n and enterotoxigenic escherichia coli f4 fimbriae |
| url | http://link.springer.com/article/10.1186/s13567-018-0519-9 |
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