The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex
The enterobacterial common antigen (ECA) is a carbohydrate polymer that is associated with the cell envelope in the Enterobacteriaceae. ECA contains a repeating trisaccharide which is polymerized by WzyE, a member of the Wzy membrane protein polymerase superfamily. WzyE activity is regulated by a me...
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| Format: | Article |
| Language: | English |
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The Royal Society
2023-03-01
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| Series: | Open Biology |
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| Online Access: | https://royalsocietypublishing.org/doi/10.1098/rsob.220373 |
| _version_ | 1797864737925496832 |
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| author | Miriam Weckener Laura S. Woodward Bradley R. Clarke Huanting Liu Philip N. Ward Audrey Le Bas David Bhella Chris Whitfield James H. Naismith |
| author_facet | Miriam Weckener Laura S. Woodward Bradley R. Clarke Huanting Liu Philip N. Ward Audrey Le Bas David Bhella Chris Whitfield James H. Naismith |
| author_sort | Miriam Weckener |
| collection | DOAJ |
| description | The enterobacterial common antigen (ECA) is a carbohydrate polymer that is associated with the cell envelope in the Enterobacteriaceae. ECA contains a repeating trisaccharide which is polymerized by WzyE, a member of the Wzy membrane protein polymerase superfamily. WzyE activity is regulated by a membrane protein polysaccharide co-polymerase, WzzE. Förster resonance energy transfer experiments demonstrate that WzyE and WzzE from Pectobacterium atrosepticum form a complex in vivo, and immunoblotting and cryo-electron microscopy (cryo-EM) analysis confirm a defined stoichiometry of approximately eight WzzE to one WzyE. Low-resolution cryo-EM reconstructions of the complex, aided by an antibody recognizing the C-terminus of WzyE, reveals WzyE sits in the central membrane lumen formed by the octameric arrangement of the transmembrane helices of WzzE. The pairing of Wzy and Wzz is found in polymerization systems for other bacterial polymers, including lipopolysaccharide O-antigens and capsular polysaccharides. The data provide new structural insight into a conserved mechanism for regulating polysaccharide chain length in bacteria. |
| first_indexed | 2024-04-09T22:58:04Z |
| format | Article |
| id | doaj.art-19c5542dd1c9491aa5e65f1ae1097f02 |
| institution | Directory Open Access Journal |
| issn | 2046-2441 |
| language | English |
| last_indexed | 2024-04-09T22:58:04Z |
| publishDate | 2023-03-01 |
| publisher | The Royal Society |
| record_format | Article |
| series | Open Biology |
| spelling | doaj.art-19c5542dd1c9491aa5e65f1ae1097f022023-03-22T11:08:53ZengThe Royal SocietyOpen Biology2046-24412023-03-0113310.1098/rsob.220373The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complexMiriam Weckener0Laura S. Woodward1Bradley R. Clarke2Huanting Liu3Philip N. Ward4Audrey Le Bas5David Bhella6Chris Whitfield7James H. Naismith8Structural Biology, The Rosalind Franklin Institute, Harwell Campus, Didcot OX11 0QS, UKCentre Biomedical Sciences, North Haugh, University of St Andrews, St Andrews KY16 9ST, UKDepartment of Molecular and Cellular Biology, The University of Guelph, Guelph, ON, CanadaCentre Biomedical Sciences, North Haugh, University of St Andrews, St Andrews KY16 9ST, UKStructural Biology, The Rosalind Franklin Institute, Harwell Campus, Didcot OX11 0QS, UKStructural Biology, The Rosalind Franklin Institute, Harwell Campus, Didcot OX11 0QS, UKMRC—University of Glasgow Centre for Virus Research, University of Glasgow, Sir Michael Stoker Building, Garscube Campus, 464 Bearsden Road, Glasgow G61 1Q, UKDepartment of Molecular and Cellular Biology, The University of Guelph, Guelph, ON, CanadaStructural Biology, The Rosalind Franklin Institute, Harwell Campus, Didcot OX11 0QS, UKThe enterobacterial common antigen (ECA) is a carbohydrate polymer that is associated with the cell envelope in the Enterobacteriaceae. ECA contains a repeating trisaccharide which is polymerized by WzyE, a member of the Wzy membrane protein polymerase superfamily. WzyE activity is regulated by a membrane protein polysaccharide co-polymerase, WzzE. Förster resonance energy transfer experiments demonstrate that WzyE and WzzE from Pectobacterium atrosepticum form a complex in vivo, and immunoblotting and cryo-electron microscopy (cryo-EM) analysis confirm a defined stoichiometry of approximately eight WzzE to one WzyE. Low-resolution cryo-EM reconstructions of the complex, aided by an antibody recognizing the C-terminus of WzyE, reveals WzyE sits in the central membrane lumen formed by the octameric arrangement of the transmembrane helices of WzzE. The pairing of Wzy and Wzz is found in polymerization systems for other bacterial polymers, including lipopolysaccharide O-antigens and capsular polysaccharides. The data provide new structural insight into a conserved mechanism for regulating polysaccharide chain length in bacteria.https://royalsocietypublishing.org/doi/10.1098/rsob.220373lipidoligosaccharidespolymeraseWzyregulating |
| spellingShingle | Miriam Weckener Laura S. Woodward Bradley R. Clarke Huanting Liu Philip N. Ward Audrey Le Bas David Bhella Chris Whitfield James H. Naismith The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex Open Biology lipid oligosaccharides polymerase Wzy regulating |
| title | The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex |
| title_full | The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex |
| title_fullStr | The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex |
| title_full_unstemmed | The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex |
| title_short | The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex |
| title_sort | lipid linked oligosaccharide polymerase wzy and its regulating co polymerase wzz from enterobacterial common antigen biosynthesis form a complex |
| topic | lipid oligosaccharides polymerase Wzy regulating |
| url | https://royalsocietypublishing.org/doi/10.1098/rsob.220373 |
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