Topology of the Shigella flexneri Enterobacterial Common Antigen polymerase WzyE

Maczuga, Nicholas; Tran, Elizabeth Ngoc Hoa; Morona, Renato

doi:10.1099/mic.0.001183

Cited by 3 publications

(2 citation statements)

References 47 publications

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“…Once flipped, ECA repeat units are polymerized by transfer of the growing chain from its Und-PP carrier to the non-reducing terminus of an incoming Und-PP-linked repeat unit. This reaction is catalysed by the integral membrane polymerase WzyE, which belongs to a larger shape, elongation, division and sporulation (SEDS) glycosyltransferase family [ 11 ]. After polymerization, the fate of the final polysaccharide polymer depends on the pathway.…”

Section: Introductionmentioning

confidence: 99%

The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex

et al. 2023

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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.

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Section: Introductionmentioning

confidence: 99%

The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex

et al. 2023

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“…The translocation of Und‐P to the periplasmic side of the IM by UppP remains a speculative. Figure adapted from Maczuga et al ( 2022a )…”

Section: Introductionmentioning

confidence: 99%

Subcellular localization of the enterobacterial common antigen GT‐E‐like glycosyltransferase, WecG

Maczuga

Tran

Morona

2022

Molecular Microbiology

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ECA has been shown to play major roles in the homeostasis of cells as mutants along its biosynthetic pathway, particularly its glycosyltransferases have been shown to induce pleiotropic phenotypes as well as induce cell wall stress pathways (Castelli & Vescovi, 2011;Jorgenson et al., 2016). Due to this, research into the roles that the ECA glycosyltransferases play in these pleiotropic phenotypes has dominated ECA research with little research being performed in understanding the enzymes themselves. WecG research has not progressed since 1988 when it was first identified by Barr et al. who described WecG as a membrane protein. Here, we show that WecG is a peripheral membrane protein, maintained to the IM via interactions facilitated by its C-terminal tail. We describe a new model of how WecG is maintained to the IM and present WecG as the second protein in the novel glycosyltransferase-fold family, GT-E. | INTRODUC TI ONEnterobacterales, an order of bacteria belonging to Gram-negatives, have developed unique adaptations which allow them to persist and

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Identification and analysis of immunoreactive proteins of Shigella flexneri in human sera and stool specimens

Singh,

Salleh,

Ahmed

et al. 2024

PeerJ

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Background The method currently available to diagnose shigellosis is insensitive and has many limitations. Thus, this study was designed to identify specific antigenic protein(s) among the cell surface associated proteins (SAPs) of Shigella that would be valuable in the development of an alternative diagnostic assay for shigellosis, particularly one that could be run using a stool sample rather than serum. Methods The SAPs of clinical isolates of S. dysenteriae, S. boydii, Shigella flexneri, and S. sonnei were extracted from an overnight culture grown at 37 °C using acidified-glycine extraction methods. Protein profiles were observed by SDS-PAGE. To determine if antibodies specific to certain Shigella SAPs were present in both sera and stool suspensions, Western blot analysis was used to detect the presence of IgA, IgG, and IgM. Results Immunoblot analysis revealed that sera from patients infected with S. flexneri recognized 31 proteins. These SAP antigens are recognized by the host humoral response during Shigella infection. Specific antibodies against these antigens were also observed in intestinal secretions of shigellosis patients. Of these 31 S. flexneri proteins, the 35 kDa protein specifically reacted against IgA present in patients’ stool suspensions. Further study illustrated the immunoreactivity of this protein in S. dysenteriae, S. boydii, and S. sonnei. This is the first report that demonstrates the presence of immunoreactive Shigella SAPs in stool suspensions. The SAPSs could be very useful in developing a simple and rapid serodiagnostic assay for shigellosis directly from stool specimens.

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Topology of the Shigella flexneri Enterobacterial Common Antigen polymerase WzyE

Cited by 3 publications

References 47 publications

The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex

The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex

Subcellular localization of the enterobacterial common antigen GT‐E‐like glycosyltransferase, WecG

Identification and analysis of immunoreactive proteins of Shigella flexneri in human sera and stool specimens

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