keyword
https://read.qxmd.com/read/36944376/the-lipid-linked-oligosaccharide-polymerase-wzy-and-its-regulating-co-polymerase-wzz-from-enterobacterial-common-antigen-biosynthesis-form-a-complex
#1
JOURNAL ARTICLE
Miriam Weckener, Laura S Woodward, Bradley R Clarke, Huanting Liu, Philip N Ward, Audrey Le Bas, David Bhella, Chris Whitfield, James H Naismith
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...
March 2023: Open Biology
https://read.qxmd.com/read/36849993/the-effect-of-o-antigen-length-determinant-wzz-on-the-immunogenicity-of-salmonella-typhimurium-for-escherichia-coli-o2-o-polysaccharides-delivery
#2
JOURNAL ARTICLE
Yue Han, Ping Luo, Huan Zeng, Pu Wang, Jiali Xu, Pengju Chen, Xindan Chen, Yuji Chen, Qiyu Cao, Ruidong Zhai, Jing Xia, Simin Deng, Anchun Cheng, Changyong Cheng, Houhui Song
Attenuated Salmonella Typhimurium is a promising antigen delivery system for live vaccines such as polysaccharides. The length of polysaccharides is a well-known key factor in modulating the immune response induced by glycoconjugates. However, the relationship between the length of Lipopolysaccharide (LPS) O-antigen (OAg) and the immunogenicity of S. Typhimurium remains unclear. In this study, we assessed the effect of OAg length determined by wzzST on Salmonella colonization, cell membrane permeability, antimicrobial activity, and immunogenicity by comparing the S...
February 27, 2023: Veterinary Research
https://read.qxmd.com/read/36622162/repeat-unit-elongations-to-produce-bacterial-complex-long-polysaccharide-chains-an-o-antigen-perspective
#3
REVIEW
Yaoqin Hong, Dalong Hu, Anthony D Verderosa, Jilong Qin, Makrina Totsika, Peter R Reeves
The O-antigen, a long polysaccharide that constitutes the distal part of the outer membrane-anchored lipopolysaccharide, is one of the critical components in the protective outer membrane of Gram-negative bacteria. Most species produce one of the structurally diverse O-antigens, with nearly all the polysaccharide components having complex structures made by the Wzx/Wzy pathway. This pathway produces repeat-units of mostly 3-8 sugars on the cytosolic face of the cytoplasmic membrane that is translocated by Wzx flippase to the periplasmic face and polymerized by Wzy polymerase to give long-chain polysaccharides...
January 9, 2023: EcoSal Plus
https://read.qxmd.com/read/35980183/identification-of-the-shigella-flexneri-wzy-domain-modulating-wzz-phs-2-interaction-and-detection-of-the-wzy-wzz-oag-complex
#4
JOURNAL ARTICLE
Alice Ascari, Elizabeth Ngoc Hoa Tran, Bart A Eijkelkamp, Renato Morona
Shigella flexneri implements the Wzy-dependent pathway to biosynthesize the O antigen (Oag) component of its surface lipopolysaccharide. The inner membrane polymerase WzySF catalyzes the repeat addition of undecaprenol-diphosphate-linked Oag (Und-PP-RUs) to produce a polysaccharide, the length of which is tightly regulated by two competing copolymerase proteins, WzzSF (short-type Oag; 10 to 17 RUs) and WzzpHS-2 (very-long-type Oag; >90 RUs). The nature of the interaction between WzySF and WzzSF /WzzpHS-2 in Oag polymerization remains poorly characterized, with the majority of the literature characterizing the individual protein constituents of the Wzy-dependent pathway...
August 18, 2022: Journal of Bacteriology
https://read.qxmd.com/read/35470793/topology-of-the-shigella-flexneri-enterobacterial-common-antigen-polymerase-wzye
#5
JOURNAL ARTICLE
Nicholas T Maczuga, Elizabeth N H Tran, Renato Morona
Enterobacteriales have evolved a specialized outer membrane polysaccharide [Enterobacterial Common Antigen (ECA)] which allows them to persist in various environmental niches. Biosynthesis of ECA initiates on the cytoplasmic leaflet of the inner membrane (IM) where glycosyltransferases assemble ECA repeat units (RUs). Complete RUs are then translocated across the IM and assembled into polymers by ECA-specific homologues of the Wzy-dependent pathway. Consisting of the membrane proteins Wzx, Wzy and Wzz, the Wzy-dependent pathway is the most common polysaccharide biosynthetic pathway in Gram-negative bacteria where it is most notably involved in LPS O antigen (Oag) biosynthesis...
April 2022: Microbiology
https://read.qxmd.com/read/34491798/identification-of-a-region-in-shigella-flexneri-wzyb-disrupting-the-interaction-with-wzz-phs2
#6
JOURNAL ARTICLE
Vincenzo Leo, Min Yan Teh, Elizabeth N H Tran, Renato Morona
Shigella flexneri can synthesise polysaccharide chains having complex sugars and a regulated number of repeating units. S. flexneri lipopolysaccharide O antigen (Oag) is synthesised by the Wzy-dependent pathway which is the most common pathway used in bacteria for polysaccharide synthesis. The inner membrane protein WzyB polymerizes the Oag repeat units into chains, while the polysaccharide co-polymerases WzzB and WzzpHS2 determine the average number of repeat units or "the modal length", termed short-type and very long-type...
September 7, 2021: Journal of Bacteriology
https://read.qxmd.com/read/34248085/physiological-and-genomic-analysis-of-newly-isolated-polysaccharide-synthesizing-cyanobacterium-chroococcus-sp-fpu101-and-chemical-analysis-of-the-exopolysaccharide
#7
JOURNAL ARTICLE
Shinya Yoshikawa, Yu Kanesaki, Akira Uemura, Kazumasa Yamada, Maiko Okajima, Tatsuo Kaneko, Kaori Ohki
A unicellular cyanobacterium that produces a large amount of exopolysaccharide (EPS) was isolated. The isolate, named Chroococcus sp. FPU101, grew between 20 and 30°C and at light intensities between 10 and 80 μmol m-2 s-1 . Purified EPS from Chroococcus sp. FPU101 had a molecular size of 5.9 × 103 kDa and contained galactose, rhamnose, fucose, xylose, mannose, glucose, galacturonic acid, and glucuronic acid at a molar ratio of 17.2:15.9:14.1:11.0:9.6:9.5:13.0:9.7. The EPS content significantly increased when the NaCl concentration in the medium was increased from 1...
July 10, 2021: Journal of General and Applied Microbiology
https://read.qxmd.com/read/33446644/structure-of-a-full-length-bacterial-polysaccharide-co-polymerase
#8
JOURNAL ARTICLE
Benjamin Wiseman, Ram Gopal Nitharwal, Göran Widmalm, Martin Högbom
Lipopolysaccharides are important components of the bacterial cell envelope that among other things act as a protective barrier against the environment and toxic molecules such as antibiotics. One of the most widely disseminated pathways of polysaccharide biosynthesis is the inner membrane bound Wzy-dependent pathway. Here we present the 3.0 Å structure of the co-polymerase component of this pathway, WzzB from E. coli solved by single-particle cryo-electron microscopy. The overall architecture is octameric and resembles a box jellyfish containing a large bell-shaped periplasmic domain with the 2-helix transmembrane domain from each protomer, positioned 32 Å apart, encircling a large empty transmembrane chamber...
January 14, 2021: Nature Communications
https://read.qxmd.com/read/33361196/polysaccharide-co-polymerase-wzzb-wzze-chimeras-reveal-transmembrane-2-region-of-wzzb-is-important-for-interaction-with-wzyb
#9
JOURNAL ARTICLE
Vincenzo Leo, Elizabeth Tran, Renato Morona
The ability of bacteria to synthesise complex polysaccharide chains at a controlled number of repeating units has wide implications for a range of biological activities that include: symbiosis, biofilm formation and immune system avoidance. Complex polysaccharide chains such as the O antigen (Oag) component of lipopolysaccharide and the enterobacterial common antigen (ECA) are synthesised by the most common polysaccharide synthesis pathway used in bacteria, known as the Wzy-dependent pathway. The Oag and ECA are polymerized into chains via the inner membrane proteins WzyB and WzyE, respectively, while the respective co-polymerases WzzB and WzzE modulate the number of repeat units in the chains or "the modal length" of the polysaccharide via a hypothesised interaction...
December 23, 2020: Journal of Bacteriology
https://read.qxmd.com/read/33075674/structural-analysis-of-the-o-antigen-polysaccharide-from-escherichia-coli-o188
#10
JOURNAL ARTICLE
Axel Furevi, Jonas Ståhle, Claudio Muheim, Spyridon Gkotzis, Klas I Udekwu, Daniel O Daley, Göran Widmalm
The structure of the O-antigen from Escherichia coli reference strain O188 (E. coli O188:H10) has been investigated. The lipopolysaccharide shows a typical nonrandom modal chain-length distribution and the sugar and absolute configuration analysis revealed d-Man, d-Glc, d-GlcN and d-GlcA as major components. The structure of the O-specific polysaccharide was determined using one- and two-dimensional 1 H and 13 C NMR spectroscopy experiments, where inter-residue correlations were identified by 1 H,13 C-heteronuclear multiple-bond correlation and 1 H,1 H-NOESY experiments, which revealed that it consists of pentasaccharide repeating units with the following structure: Biosynthetic aspects and NMR analysis are consistent with the presented structure as the biological repeating unit...
December 2020: Carbohydrate Research
https://read.qxmd.com/read/32411015/physiological-characteristics-and-environment-adaptability-of-reef-building-corals-at-the-wuzhizhou-island-of-south-china-sea
#11
JOURNAL ARTICLE
Huili Xu, Boxuan Feng, Minrui Xie, Yuxiao Ren, Jingquan Xia, Yu Zhang, Aimin Wang, Xiubao Li
The health of coral reef has declined significantly around the world due to the impact of human activities and natural environment changes, and corals have to develop effective resistance mechanisms to survive. In this study, we examined the physiological characteristics and Symbiodiniaceae types of four dominant scleractinian corals in the reefs at the Wuzhizhou Island (WZZ) in South China Sea. The water environmental conditions are complex on the north side of WZZ due to regional geography and tourism development, and all corals had their unique physiological conditions and Symbiodiniaceae types...
2020: Frontiers in Physiology
https://read.qxmd.com/read/31109993/unique-regions-of-the-polysaccharide-co-polymerase-wzz-2-from-pseudomonas-aeruginosa-are-essential-for-o-specific-antigen-chain-length-control
#12
JOURNAL ARTICLE
Steven M Huszczynski, Chelsea Coumoundouros, Phi Pham, Joseph S Lam, Cezar M Khursigara
The outer leaflet of the outer membrane of nearly all Gram-negative bacteria contains lipopolysaccharide (LPS). The distal end of LPS may be capped with O antigen, a long polysaccharide that can range from a few to hundreds of sugars in length. The chain length of the polysaccharide has many implications for bacterial survival and, consequently, is tightly controlled. In the Wzx/Wzy-dependent route of O antigen synthesis, one or more Wzz proteins determine the chain length via an unknown mechanism. To gain insight into this mechanism, we identified and characterized important regions of two Wzz proteins in Pseudomonas aeruginosa serotype O13, which confer the production of "long" (Wzz1 ) and "very long" (Wzz2 ) chain lengths, respectively...
May 20, 2019: Journal of Bacteriology
https://read.qxmd.com/read/30872471/glycoconjugate-vaccine-using-a-genetically-modified-o-antigen-induces-protective-antibodies-to-francisella-tularensis
#13
JOURNAL ARTICLE
Giuseppe Stefanetti, Nihal Okan, Avner Fink, Erica Gardner, Dennis L Kasper
Francisella tularensis is the causative agent of tularemia, a category A bioterrorism agent. The lipopolysaccharide (LPS) O antigen (OAg) of F. tularensis has been considered for use in a glycoconjugate vaccine, but conjugate vaccines tested so far have failed to confer protection necessary against aerosolized pulmonary bacterial challenge. When F. tularensis OAg was purified under standard conditions, the antigen had a small molecular size [25 kDa, low molecular weight (LMW)]. Using milder extraction conditions, we found the native OAg had a larger molecular size [80 kDa, high molecular weight (HMW)], and in a mouse model of tularemia, a glycoconjugate vaccine made with the HMW polysaccharide coupled to tetanus toxoid (HMW-TT) conferred better protection against intranasal challenge than a conjugate made with the LMW polysaccharide (LMW-TT)...
March 14, 2019: Proceedings of the National Academy of Sciences of the United States of America
https://read.qxmd.com/read/30615987/development-and-potential-use-of-an-edwardsiella-ictaluri-wzz-mutant-as-a-live-attenuated-vaccine-against-enteric-septicemia-in-pangasius-hypophthalmus-tra-catfish
#14
JOURNAL ARTICLE
Tran H Triet, Bui T T Tinh, Le V Hau, Tran V Huong, Nguyen-Quoc Binh
Edwardsiella ictaluri is a causative agent of enteric septicemia of catfish (ESC), a seriously lethal disease in Vietnamese catfish (Pangasius hypophthalmus). A safe and effective vaccine against ESC is currently an urgent demand due to antibiotic overuse in pangasius farms has led to an alarming antimicrobial resistance. In this study, two E. ictaluri wzzE mutants (WzM-L3, deficient in a 1038bp-entire wzzE gene and WzM-S3, a 245bp-partial deletion of wzzE) were developed and their protection efficiacy was evaluated in hatched pangasius against ESC by immersion vaccination...
January 4, 2019: Fish & Shellfish Immunology
https://read.qxmd.com/read/29998926/physiological-properties-and-genetic-analysis-related-to-exopolysaccharide-eps-production-in-the-fresh-water-unicellular-cyanobacterium-aphanothece-sacrum-suizenji-nori
#15
JOURNAL ARTICLE
Kaori Ohki, Yu Kanesaki, Noriyuki Suzuki, Maiko Okajima, Tatsuo Kaneko, Shinya Yoshikawa
The clonal strains, phycoerythrin(PE)-rich- and PE-poor strains, of the unicellular, fresh water cyanobacterium Aphanothece sacrum (Suringar) Okada (Suizenji Nori, in Japanese) were isolated from traditional open-air aquafarms in Japan. A. sacrum appeared to be oligotrophic on the basis of its growth characteristics. The optimum temperature for growth was around 20°C. Maximum growth and biomass increase at 20°C was obtained under light intensities between 40 to 80 μmol m-2 s-1 (fluorescent lamps, 12 h light/12 h dark cycles) and between 40 to 120 μmol m-2 s-1 for PE-rich and PE-poor strains, respectively, of A...
July 12, 2018: Journal of General and Applied Microbiology
https://read.qxmd.com/read/29996532/possible-role-of-envelope-components-in-the-extreme-copper-resistance-of-the-biomining-acidithiobacillus-ferrooxidans
#16
JOURNAL ARTICLE
Nia Oetiker, Rodrigo Norambuena, Cristóbal Martínez-Bussenius, Claudio A Navarro, Fernando Amaya, Sergio A Álvarez, Alberto Paradela, Carlos A Jerez
Acidithiobacillus ferrooxidans resists extremely high concentrations of copper. Strain ATCC 53993 is much more resistant to the metal compared with strain ATCC 23270, possibly due to the presence of a genomic island in the former one. The global response of strain ATCC 53993 to copper was analyzed using iTRAQ (isobaric tag for relative and absolute quantitation) quantitative proteomics. Sixty-seven proteins changed their levels of synthesis in the presence of the metal. On addition of CusCBA efflux system proteins, increased levels of other envelope proteins, such as a putative periplasmic glucan biosynthesis protein (MdoG) involved in the osmoregulated synthesis of glucans and a putative antigen O polymerase (Wzy), were seen in the presence of copper...
July 10, 2018: Genes
https://read.qxmd.com/read/29866904/novel-role-of-visp-and-the-wzz-system-during-o-antigen-assembly-in-salmonella-enterica-serovar-typhimurium-pathogenesis
#17
JOURNAL ARTICLE
Patrick da Silva, Fernanda Z Manieri, Carmen M Herrera, M Stephen Trent, Cristiano G Moreira
Salmonella enterica serovars are associated with diarrhea and gastroenteritis and are a helpful model for understanding host-pathogen mechanisms. Salmonella enterica serovar Typhimurium regulates the distribution of O antigen (OAg) and presents a trimodal distribution based on Wzy polymerase and the WzzST (long-chain-length OAg [L-OAg]) and WzzfepE (very-long-chain-length OAg [VL-OAg]) copolymerases; however, several mechanisms regulating this process remain unclear. Here, we report that LPS modifications modulate the infectious process and that OAg chain length determination plays an essential role during infection...
August 2018: Infection and Immunity
https://read.qxmd.com/read/29603538/overexpression-of-o-polysaccharide-chain-length-regulators-in-gram-negative-bacteria-using-the-wzx-wzy-dependent-pathway-enhances-production-of-defined-modal-length-o-polysaccharide-polymers-for-use-as-haptens-in-glycoconjugate-vaccines
#18
JOURNAL ARTICLE
N Hegerle, J Bose, G Ramachandran, J E Galen, M M Levine, R Simon, S M Tennant
AIMS: O-polysaccharide (OPS) molecules are protective antigens for several bacterial pathogens, and have broad utility as components of glycoconjugate vaccines. Variability in the OPS chain length is one obstacle towards further development of these vaccines. Introduction of sizing steps during purification of OPS molecules of suboptimal or of mixed lengths introduces additional costs and complexity while decreasing the final yield. The overall goal of this study was to demonstrate the utility of engineering Gram-negative bacteria to produce homogenous O-polysaccharide populations that can be used as the basis of carbohydrate vaccines by overexpressing O-polysaccharide chain length regulators of the Wzx-/Wzy-dependent pathway...
August 2018: Journal of Applied Microbiology
https://read.qxmd.com/read/28883503/differential-impact-of-lipopolysaccharide-defects-caused-by-loss-of-rfah-in-yersinia-pseudotuberculosis-and-yersinia-pestis
#19
JOURNAL ARTICLE
Jared M Hoffman, Shea Sullivan, Erin Wu, Eric Wilson, David L Erickson
RfaH enhances transcription of a select group of operons controlling bacterial surface features such as lipopolysaccharide (LPS). Previous studies have suggested that rfaH may be required for Yersinia pseudotuberculosis resistance to antimicrobial chemokines and survival during mouse infections. In order to further investigate the role of RfaH in LPS synthesis, resistance to host defense peptides, and virulence of Yersinia, we constructed ΔrfaH mutants of Y. pseudotuberculosis IP32953 and Y. pestis KIM6+. Loss of rfaH affected LPS synthesis in both species, resulting in a shorter core oligosaccharide...
September 7, 2017: Scientific Reports
https://read.qxmd.com/read/28434914/full-length-oligomeric-structure-of-wzz-determined-by-cryoelectron-microscopy-reveals-insights-into-membrane-bound-states
#20
JOURNAL ARTICLE
Richard F Collins, Vasileios Kargas, Brad R Clarke, C Alistair Siebert, Daniel K Clare, Peter J Bond, Chris Whitfield, Robert C Ford
Wzz is an integral inner membrane protein involved in regulating the length of lipopolysaccharide O-antigen glycans and essential for the virulence of many Gram-negative pathogens. In all Wzz homologs, the large periplasmic domain is proposed to be anchored by two transmembrane helices, but no information is available for the transmembrane and cytosolic domains. Here we have studied purified oligomeric Wzz complexes using cryoelectron microscopy and resolved the transmembrane regions within a semi-continuous detergent micelle...
May 2, 2017: Structure
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