Structure of the N-Linked Glycan Present on Multiple Glycoproteins in the Gram-negative Bacterium, Campylobacter jejuni

Young, N. Martin; Brisson, Jean‐Robert; Kelly, John F.; Watson, David; Tessier, Luc; Lanthier, Patricia; Jarrell, Harold C.; Cadotte, Nicolas; Michael, Frank St.; Aberg, Erika; Szymanski, Christine M.

doi:10.1074/jbc.m206114200

Cited by 397 publications

(418 citation statements)

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“…NIH-PA Author Manuscript NIH-PA Author Manuscript (7)(8)(9)(10). The genes coding for enzymes that synthesize these glycan moieties have been found in clusters throughout the C. jejuni genome.…”

Section: Nih-pa Author Manuscriptmentioning

confidence: 99%

“…Pseudaminic acid, the major modification of flagellin, is synthesized via the UDP-GlcNAc dehydratase PseB pathway (30,18). GalNAc, a major component in the biosynthesis of glycoprotein N-linked heptasaccharide, lipooligosaccharide (LOS), and capsule (7,8,10), is synthesized intracellularly by the C4 epimerase Gne (31). Table 3 lists the kinetic parameters of PseB and Gne derived from other studies, as well as GST-PglF from this study.…”

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confidence: 99%

“…A common N-linked glycan has been detected in at least 22 periplasmic and cell-surface proteins (14,10) in C. jejuni, and the Pgl pathway has been identified as the source of its biosynthesis. Genes of the pgl locus in the pathogen have been designated cj1119-cj1130 and the structure of the N-linked glycan has been shown to be the heptasaccharide GalNAc-α1,4-GalNAc-α1,4-(Glcβ1,3)-GalNAc-α1,4-GalNAc-α1,4-GalNAc-α1,3-Bac2,4diNAc-_1-Asn where GalNAc, GlcNAc, and Glc represent N-acetylgalactosamine, N-acetylglucosamine, and glucose, respectively, ( Figure 1) (10).…”

mentioning

confidence: 99%

“…Genes of the pgl locus in the pathogen have been designated cj1119-cj1130 and the structure of the N-linked glycan has been shown to be the heptasaccharide GalNAc-α1,4-GalNAc-α1,4-(Glcβ1,3)-GalNAc-α1,4-GalNAc-α1,4-GalNAc-α1,3-Bac2,4diNAc-_1-Asn where GalNAc, GlcNAc, and Glc represent N-acetylgalactosamine, N-acetylglucosamine, and glucose, respectively, ( Figure 1) (10). Assembly of this bacterial glycan begins with pyrophosphate bond formation between the Bac2,4diNAc phosphate and undecaprenylphosphate (Und-P) by Cj1124c (PglC) to form Bac2,4diNAc-α1-PP-Und (15).…”

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In Vitro Biosynthesis of UDP-N,N‘-Diacetylbacillosamine by Enzymes of the Campylobacter jejuni General Protein Glycosylation System

et al. 2006

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In Campylobacter jejuni the sugar 2,4-diacetamido-2,4,6-trideoxy-α-D-glucopyranose, termed N,N ′-diacetylbacillosamine (Bac2,4diNAc), is the first carbohydrate in the glycoprotein N-linked heptasaccharide. Starting with uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc) two enzymes of the general protein glycosylation (Pgl) pathway in C. jejuni (PglF and PglE) have been recently shown to modify this sugar nucleotide to form UDP-2-acetamido-4-amino-2,4,6-trideoxy-α-D-glycopyranose (UDP-4-amino-sugar) [Schoenhofen, I. C., et al. (2006) J Biol Chem 281, 723-732]. PglD has been proposed to catalyze the final step in N,N′-diacetylbacillosamine synthesis by N-acetylation of the UDP-4-amino-sugar at the C4 position. We have cloned, overexpressed and purified PglD from the pgl locus of C. jejuni NCTC 11168 and identified it as the acetyltransferase that modifies the UDP-4-amino-sugar to form UDP-N,N′-diacetylbacillosamine, utilizing acetyl coenzyme A as the acetyl group donor. The UDP-N,N′-diacetylbacillosamine product was purified from the reaction by reverse phase C18 HPLC and the structure determined by NMR analysis. Additionally, the full-length PglF was overexpressed and purified in the presence of detergent as a GST-fusion protein allowing for derivation of kinetic parameters. We found that the UDP-4-aminosugar was readily synthesized from UDP-GlcNAc in a coupled reaction using PglF and PglE. We also demonstrate the in vitro biosynthesis of the complete heptasaccharide lipid-linked donor by coupling the action of eight enzymes (PglF, PglE, PglD, PglC, PglA, PglJ, PglH, and PglI) in the Pgl pathway in a single reaction vessel.Campylobacter jejuni is the Gram-negative enteropathogen identified as the primary cause of gastroenteritis in humans and the most frequent infection to precede the peripheral neuropathy Guillain-Barré syndrome (1-3). Infection of humans generally occurs by ingestion of contaminated livestock or water. Although the mechanism of infection is not clearly understood, production of glycolipids and glycoproteins by the pathogen have been found to influence cell motility, host-cell interactions, and competence for DNA uptake (4-6). As resistance to antimicrobial agents rises, the potential for development of novel therapeutics against enzymes that produce glycoconjugates has intensified efforts targeted at the characterization of their biosynthetic pathways. In C. jejuni four major glycan structures have been identified: lipooligosaccharide (LOS), capsule, O-linked glycan, and N-linked glycan *To whom correspondence should be addressed: Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA 02139. E-mail: imper@mit.edu, (v) 617-253-1838, (f) NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript (7)(8)(9)(10). The genes coding for enzymes that synthesize these glycan moieties have been found in clusters throughout the C. jejuni genome. The respective biochemical functions of the gene products have been assigned on the b...

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Section: Nih-pa Author Manuscriptmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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In Vitro Biosynthesis of UDP-N,N‘-Diacetylbacillosamine by Enzymes of the Campylobacter jejuni General Protein Glycosylation System

et al. 2006

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“…[21,22] Based on the availability of molecular tools, the Pgl protein N-glycosylation system from Campylobacter jejuni [23] and the E. coli O7 antigen biosynthesis system [24] were used for the design of SgsE-neoglycoproteins. The C. jejuni Pgl enzymes synthesize a heptasaccharide with the structure D-GalNAc-α1,4-D-GalNAc-α1,4-(D-Glc-β1,3)-D-GalNAc-α1,4-D-GalNAc-α1,4-D-GalNAc-α1,3-D-Bac, where Bac is 2,4-diacetamido-2,4,6-trideoxy-D-Glc, [25] involving the lipid carrier undecaprenylpyrophosphate. [26] The heptasaccharide is transferred by the PglB oligosaccharyl:protein transferase to asparagine residues present in the target protein consensus sequence D-X-N-Z-S/T (where X and Z are any amino acid except proline).…”

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confidence: 99%

Recombinant Glycans on an S‐Layer Self‐Assembly Protein: A New Dimension for Nanopatterned Biomaterials

Steiner

Hanreich

Kainz

et al. 2008

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Crucial biological phenomena are mediated through carbohydrates that are displayed in a defined manner and interact with molecular scale precision. We lay the groundwork for the integration of recombinant carbohydrates into a "biomolecular construction kit" for the design of new biomaterials, by utilizing the self-assembly system of the crystalline cell surface (S)-layer protein SgsE of Geobacillus stearothermophilus NRS 2004/3a. SgsE is a naturally O-glycosylated protein, with intrinsic properties that allow it to function as a nanopatterned matrix for the periodic display of glycans. By using a combined carbohydrate/protein engineering approach, two types of S-layer neoglycoproteins are produced in Escherichia coli. Based on the identification of a suitable periplasmic targeting system for the SgsE self-assembly protein as a cellular prerequisite for Europe PMC Funders Author ManuscriptsEurope PMC Funders Author Manuscripts protein glycosylation, and on engineering of one of the natural protein O-glycosylation sites into a target for N-glycosylation, the heptasaccharide from the AcrA protein of Campylobacter jejuni and the O7 polysaccharide of E. coli are co-or post-translationally transferred to the S-layer protein by the action of the oligosaccharyltransferase PglB. The degree of glycosylation of the Slayer neoglycoproteins after purification from the periplasmic fraction reaches completeness. Electron microscopy reveals that recombinant glycosylation is fully compatible with the S-layer protein self-assembly system. Tailor-made ("functional") nanopatterned, self-assembling neoglycoproteins may open up new strategies for influencing and controlling complex biological systems with potential applications in the areas of biomimetics, drug targeting, vaccine design, or diagnostics.

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Comparative genomics of the ɛ‐proteobacterial human pathogens H elicobacter pylori and C ampylobacter jejuni

Dorrell¹,

Brendan²

2005

Encyclopedia of Genetics, Genomics, Proteomics and Bioinformatics

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Helicobacter pylori and Campylobacter jejuni are among the most common bacterial pathogens encountered by humans, but despite the prevalence and importance of the diseases caused by these organisms, they have only been recognized as human pathogens within the last 25 years. However, they are now among the most comprehensively studied organisms, due in no small part to the availability of complete genome sequences for both bacteria. Subsequent genomic studies have started to explain the genetic, ecological, and pathogenic similarities and differences between these two major human pathogens. Understanding the dynamic and complex interplay between these pathogens and the host will place the emphasis on converting this knowledge into appropriate intervention strategies to reduce the burden on human health.

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Structure of the N-Linked Glycan Present on Multiple Glycoproteins in the Gram-negative Bacterium, Campylobacter jejuni

Cited by 397 publications

References 47 publications

In Vitro Biosynthesis of UDP-N,N‘-Diacetylbacillosamine by Enzymes of the Campylobacter jejuni General Protein Glycosylation System

In Vitro Biosynthesis of UDP-N,N‘-Diacetylbacillosamine by Enzymes of the Campylobacter jejuni General Protein Glycosylation System

Recombinant Glycans on an S‐Layer Self‐Assembly Protein: A New Dimension for Nanopatterned Biomaterials

Comparative genomics of the ɛ‐proteobacterial human pathogens H elicobacter pylori and C ampylobacter jejuni

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