Identification of the capsular polysaccharides of Type D and F Pasteurella multocida as unmodified heparin and chondroitin, respectively

DeAngelis, Paul L.; Gunay, Nur Sibel; Toida, Toshihiko; Mao, Wenjun; Linhardt, Robert J.

doi:10.1016/s0008-6215(02)00219-7

Cited by 101 publications

(75 citation statements)

References 15 publications

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“…Uniformly labeled 15 N-HA polysaccharide was produced by fermentation (3 days at 30°C) of E. coli K5 transfected with recombinant HA synthase from Pasteurella multocida (24) in M9 minimal media with 15 NH 4 Cl (Ͼ99 atom %; Spectra Stable Isotopes) as the nitrogen source. The polymer in the media was purified by cetylpyridinium chloride precipitation, DNase/RNase treatment, chloroform extraction, and reverse phase extraction as described previously (25) Data Processing and Structure Calculations-Data were processed using FELIX 2.3 (Biosym Inc.) and referenced and analyzed with XEasy as described previously (11). NOE intensities for each data set were calibrated using interproton distances in regions of secondary structure and converted into three distance restraint categories with limits of 2.7, 3.5, and 5.0 Å.…”

Section: Methodsmentioning

confidence: 99%

The Link Module from Ovulation- and Inflammation-associated Protein TSG-6 Changes Conformation on Hyaluronan Binding

Blundell

Mahoney²,

Almond

et al. 2003

Journal of Biological Chemistry

256

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The solution structure of the Link module from human TSG-6, a hyaladherin with important roles in inflammation and ovulation, has been determined in both its free and hyaluronan-bound conformations. This reveals a well defined hyaluronan-binding groove on one face of the Link module that is closed in the absence of ligand. The groove is lined with amino acids that have been implicated in mediating the interaction with hyaluronan, including two tyrosine residues that appear to form essential intermolecular hydrogen bonds and two basic residues capable of supporting ionic interactions. This is the first structure of a non-enzymic hyaladherin in its active state, and identifies a ligand-induced conformational change that is likely to be conserved across the Link module superfamily. NMR and isothermal titration calorimetry experiments with defined oligosaccharides have allowed us to infer the minimum length of hyaluronan that can be accommodated within the binding site and its polarity in the groove; these data have been used to generate a model of the complex formed between the Link module and a hyaluronan octasaccharide.Hyaluronan (HA), 1 a high molecular weight polysaccharide with a central role in extracellular matrix organization and cell adhesion in mammals (1), is essential to a wide range of normal physiological processes including development, immunology, and reproduction (2-4). Alterations in the metabolism and localization of this molecule underlie the progression of many diseases, for instance arthritis, pulmonary/vascular disorders, and cancer (5, 6). These diverse biological activities may seem surprising for a linear polymer composed entirely of a repeating disaccharide (i.e. -glucuronic acid-␤Ϫ1,3-N-acetylglucosamine-␤Ϫ1,4-; up to 10 7 Da) that, unlike other glycosaminoglycans, is neither attached to a core protein nor sulfated. This functional complexity is thought to arise from the interaction of HA with a large number of specific HA-binding proteins (7), which can form structurally diverse complexes (see Ref. 8). The majority of these "hyaladherins" belong to a superfamily of proteins that share a common ϳ100 amino acid domain, termed a Link module, that mediates the interaction with HA.Previously we have determined the solution structure of the Link module from human TSG-6 (the protein product of the tumor necrosis factor-stimulated gene-6 (9)), thereby defining the consensus fold for this superfamily (10). In TSG-6, a 35-kDa secreted protein composed mainly of contiguous Link and CUB modules, the Link module is sufficient to mediate a high affinity interaction with HA (10, 11); this has been termed a "type A" HA-binding domain (7). The HA receptor CD44, which has an important role in mediating lymphocyte migration, however, requires N-and C-terminal extensions to its Link module for correct folding and functional activity of its type B interaction domain. Most other members of the superfamily, such as link proteins and chondroitin-sulfate proteoglycans (critical for extracellular matrix organiza...

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Section: Methodsmentioning

confidence: 99%

The Link Module from Ovulation- and Inflammation-associated Protein TSG-6 Changes Conformation on Hyaluronan Binding

Blundell

Mahoney²,

Almond

et al. 2003

Journal of Biological Chemistry

256

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“…Identical CPS structures can also be found in other bacteria. For example, α-(2→8)-linked PSA is also produced by N. meningitidis serogroup B and Pasteurella haemolytica serogroup A2 (9, 10), whereas P. multocida type D produces a nonsulfated heparosan CPS polymer (11).…”

mentioning

confidence: 99%

Conserved glycolipid termini in capsular polysaccharides synthesized by ATP-binding cassette transporter-dependent pathways in Gram-negative pathogens

Willis

Stupak

Richards

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

117

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Bacterial capsules are surface layers made of long-chain polysaccharides. They are anchored to the outer membrane of many Gram-negative bacteria, including pathogens such as Escherichia coli , Neisseria meningitidis , Haemophilus influenzae , and Pasteurella multocida . Capsules protect pathogens from host defenses including complement-mediated killing and phagocytosis and therefore represent a major virulence factor. Capsular polysaccharides are synthesized by enzymes located in the inner (cytoplasmic) membrane and are then translocated to the cell surface. Whereas the enzymes that synthesize the polysaccharides have been studied in detail, the structure and biosynthesis of the anchoring elements have not been definitively resolved. Here we determine the structure of the glycolipid attached to the reducing terminus of the polysialic acid capsular polysaccharides from E. coli K1 and N. meningitidis group B and the heparosan-like capsular polysaccharide from E. coli K5. All possess the same unique glycolipid terminus consisting of a lyso-phosphatidylglycerol moiety with a β-linked poly-(3-deoxy- d - manno -oct-2-ulosonic acid) (poly-Kdo) linker attached to the reducing terminus of the capsular polysaccharide.

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“…P. multocida Carter type A (fowl cholera and pasteurellosis pathogen) makes a hyaluronan (HA) capsule (11). The capsules of type D (swine atrophic rhinitis pathogen) and type F (minor fowl cholera pathogen) are composed of unsulfated N-acetylheparosan (heparosan) or unsulfated chondroitin, respectively (5). Dual-action synthases were identified for type A (PmHAS), D (PmHS), and F (PmCS) strains that polymerize the GAG chain disaccharide repeat by transferring both GlcUA and hexosamine (either GlcNAc or GalNAc, depending on the GAG) monosaccharides derived from uridine diphospho (UDP) precursors (3,(6)(7)(8).…”

mentioning

confidence: 99%

Identification of a Distinct, Cryptic Heparosan Synthase from Pasteurella multocida Types A, D, and F

DeAngelis

White

2004

J Bacteriol

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The extracellular polysaccharide capsules of Pasteurella multocida types A, D, and F are composed of hyaluronan, N-acetylheparosan (heparosan or unsulfated, unepimerized heparin), and unsulfated chondroitin, respectively. Previously, a type D heparosan synthase, a glycosyltransferase that forms the repeating disaccharide heparosan backbone, was identified. Here, a ϳ73% identical gene product that is encoded outside of the capsule biosynthesis locus was also shown to be a functional heparosan synthase. Unlike PmHS1, the PmHS2 enzyme was not stimulated greatly by the addition of an exogenous polymer acceptor and yielded smallermolecular-weight-product size distributions. Virtually identical hssB genes are found in most type A, D, and F isolates. The occurrence of multiple polysaccharide synthases in a single strain invokes the potential for capsular variation.

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Identification of the capsular polysaccharides of Type D and F Pasteurella multocida as unmodified heparin and chondroitin, respectively

Cited by 101 publications

References 15 publications

The Link Module from Ovulation- and Inflammation-associated Protein TSG-6 Changes Conformation on Hyaluronan Binding

The Link Module from Ovulation- and Inflammation-associated Protein TSG-6 Changes Conformation on Hyaluronan Binding

Conserved glycolipid termini in capsular polysaccharides synthesized by ATP-binding cassette transporter-dependent pathways in Gram-negative pathogens

Identification of a Distinct, Cryptic Heparosan Synthase from Pasteurella multocida Types A, D, and F

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