A structural mechanism for bacterial autotransporter glycosylation by a dodecameric heptosyltransferase family
A large group of bacterial virulence autotransporters including AIDA-I from diffusely adhering E. coli (DAEC) and TibA from enterotoxigenic E. coli (ETEC) require hyperglycosylation for functioning. Here we demonstrate that TibC from ETEC harbors a heptosyltransferase activity on TibA and AIDA-I, de...
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2014-10-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/03714 |
| _version_ | 1811201054162812928 |
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| author | Qing Yao Qiuhe Lu Xiaobo Wan Feng Song Yue Xu Mo Hu Alla Zamyatina Xiaoyun Liu Niu Huang Ping Zhu Feng Shao |
| author_facet | Qing Yao Qiuhe Lu Xiaobo Wan Feng Song Yue Xu Mo Hu Alla Zamyatina Xiaoyun Liu Niu Huang Ping Zhu Feng Shao |
| author_sort | Qing Yao |
| collection | DOAJ |
| description | A large group of bacterial virulence autotransporters including AIDA-I from diffusely adhering E. coli (DAEC) and TibA from enterotoxigenic E. coli (ETEC) require hyperglycosylation for functioning. Here we demonstrate that TibC from ETEC harbors a heptosyltransferase activity on TibA and AIDA-I, defining a large family of bacterial autotransporter heptosyltransferases (BAHTs). The crystal structure of TibC reveals a characteristic ring-shape dodecamer. The protomer features an N-terminal β-barrel, a catalytic domain, a β-hairpin thumb, and a unique iron-finger motif. The iron-finger motif contributes to back-to-back dimerization; six dimers form the ring through β-hairpin thumb-mediated hand-in-hand contact. The structure of ADP-D-glycero-β-D-manno-heptose (ADP-D,D-heptose)-bound TibC reveals a sugar transfer mechanism and also the ligand stereoselectivity determinant. Electron-cryomicroscopy analyses uncover a TibC–TibA dodecamer/hexamer assembly with two enzyme molecules binding to one TibA substrate. The complex structure also highlights a high efficient hyperglycosylation of six autotransporter substrates simultaneously by the dodecamer enzyme complex. |
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| id | doaj.art-3e052ecafd84487b8047d1b57d6e1efc |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:14:24Z |
| publishDate | 2014-10-01 |
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| spelling | doaj.art-3e052ecafd84487b8047d1b57d6e1efc2022-12-22T03:52:17ZengeLife Sciences Publications LtdeLife2050-084X2014-10-01310.7554/eLife.03714A structural mechanism for bacterial autotransporter glycosylation by a dodecameric heptosyltransferase familyQing Yao0Qiuhe Lu1Xiaobo Wan2Feng Song3Yue Xu4Mo Hu5Alla Zamyatina6Xiaoyun Liu7Niu Huang8Ping Zhu9Feng Shao10Dr Feng Shao's Laboratory, National Institute of Biological Sciences, Beijing, ChinaDr Feng Shao's Laboratory, National Institute of Biological Sciences, Beijing, ChinaDr Niu Huang's Laboratory, National Institute of Biological Sciences, Beijing, ChinaNational Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China; Shandong Provincial Key Laboratory of Functional Macromolecular Biophysics, Institute of Biophysics, Dezhou University, Dezhou, ChinaDr Feng Shao's Laboratory, National Institute of Biological Sciences, Beijing, ChinaInstitute of Analytic Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing, China; Synthetic Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, ChinaDepartment of Chemistry, University of Natural Resources and Life Sciences, Vienna, AustriaInstitute of Analytic Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing, China; Synthetic Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, ChinaDr Niu Huang's Laboratory, National Institute of Biological Sciences, Beijing, ChinaNational Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, ChinaDr Feng Shao's Laboratory, National Institute of Biological Sciences, Beijing, China; National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, ChinaA large group of bacterial virulence autotransporters including AIDA-I from diffusely adhering E. coli (DAEC) and TibA from enterotoxigenic E. coli (ETEC) require hyperglycosylation for functioning. Here we demonstrate that TibC from ETEC harbors a heptosyltransferase activity on TibA and AIDA-I, defining a large family of bacterial autotransporter heptosyltransferases (BAHTs). The crystal structure of TibC reveals a characteristic ring-shape dodecamer. The protomer features an N-terminal β-barrel, a catalytic domain, a β-hairpin thumb, and a unique iron-finger motif. The iron-finger motif contributes to back-to-back dimerization; six dimers form the ring through β-hairpin thumb-mediated hand-in-hand contact. The structure of ADP-D-glycero-β-D-manno-heptose (ADP-D,D-heptose)-bound TibC reveals a sugar transfer mechanism and also the ligand stereoselectivity determinant. Electron-cryomicroscopy analyses uncover a TibC–TibA dodecamer/hexamer assembly with two enzyme molecules binding to one TibA substrate. The complex structure also highlights a high efficient hyperglycosylation of six autotransporter substrates simultaneously by the dodecamer enzyme complex.https://elifesciences.org/articles/03714bacterial autotransporterglycosyltransferasebacterial pathogenesiscryo-EMenzyme complexenzyme catalysis |
| spellingShingle | Qing Yao Qiuhe Lu Xiaobo Wan Feng Song Yue Xu Mo Hu Alla Zamyatina Xiaoyun Liu Niu Huang Ping Zhu Feng Shao A structural mechanism for bacterial autotransporter glycosylation by a dodecameric heptosyltransferase family eLife bacterial autotransporter glycosyltransferase bacterial pathogenesis cryo-EM enzyme complex enzyme catalysis |
| title | A structural mechanism for bacterial autotransporter glycosylation by a dodecameric heptosyltransferase family |
| title_full | A structural mechanism for bacterial autotransporter glycosylation by a dodecameric heptosyltransferase family |
| title_fullStr | A structural mechanism for bacterial autotransporter glycosylation by a dodecameric heptosyltransferase family |
| title_full_unstemmed | A structural mechanism for bacterial autotransporter glycosylation by a dodecameric heptosyltransferase family |
| title_short | A structural mechanism for bacterial autotransporter glycosylation by a dodecameric heptosyltransferase family |
| title_sort | structural mechanism for bacterial autotransporter glycosylation by a dodecameric heptosyltransferase family |
| topic | bacterial autotransporter glycosyltransferase bacterial pathogenesis cryo-EM enzyme complex enzyme catalysis |
| url | https://elifesciences.org/articles/03714 |
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