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Membrane Association Dictates Ligand Specificity for the Innate Immune Receptor NOD2

The human gut must regulate its immune response to resident and pathogenic bacteria, numbering in the trillions. The peptidoglycan component of the bacterial cell wall is a dense and rigid structure that consists of polymeric carbohydrates and highly cross-linked peptides which offers protection from the host and surrounding environment. Nucleotide-binding oligomerization domain-containing protein 2 (NOD2), a human membrane-associated innate immune receptor found in the gut epithelium and mutated in an estimated 30% of Crohn’s disease patients, binds to peptidoglycan fragments and initiates an immune response. Using a combination of chemical synthesis, advanced analytical assays, and protein biochemistry, we tested the binding of a variety of synthetic peptidoglycan fragments to wild-type (WT)-NOD2. Only when the protein was presented in the native membrane did binding measurements correlate with a NOD2-dependent nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappaB) response, supporting the hypothesis that the native-membrane environment confers ligand specificity to the NOD2 receptor for NF-kappaB signaling. While N-acetyl-muramyl dipeptide (MDP) has been thought to be the minimal peptidoglycan fragment necessary to activate a NOD2-dependent immune response, we found that fragments with and without the dipeptide moiety are capable of binding and activating a NOD2-dependent NF-kappaB response, suggesting that the carbohydrate moiety of the peptidoglycan fragments is the minimal functional epitope. This work highlights the necessity of studying NOD2-ligand binding in systems that resemble the receptor’s natural environment, as the cellular membrane and/or NOD2 interacting partners appear to play a crucial role in ligand binding and in triggering an innate immune response.

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Reference£º
Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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Synthesis of derivatized chitooligomers using transglycosidases engineered from the fungal GH20 beta-N-acetylhexosaminidase

Abstract The synthesis of oligosaccharides using mutant glycosidases has been dynamically developing due to the need for novel carbohydrate-based materials. Chitooligomers (beta-1?4-linked oligomers of N-acetylglucosamine) are bioactive compounds applicable in many industrial and pharmacological areas; however, their accessibility is still rather low. In this work, GH20 beta-N-acetylhexosaminidase from the fungus Talaromyces flavus was engineered by site-directed mutagenesis to obtain three efficiently transglycosylating variants with ca. 200-times suppressed hydrolytic activity. Thus, we have prepared the first GH20 transglycosidases. In the reactions catalyzed by these mutant beta-N-acetylhexosaminidases we were able to easily prepare and isolate both natural and modified chitooligomers in sufficient amounts for their complete spectral characterization and possible further application. The presented method for the synthesis of chitooligomers with aglycones suitable for linking to other biological structures is simple and robust enough to be easily scaled up.

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Reference£º
Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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Purification, characterization, and analysis of antibacterial activity of a serum lectin from the grub of rhinoceros beetle, Oryctes rhinoceros

Lectins, also recognized as hemagglutinins, are multivalent proteins, and their desirable quality of fine sugar-binding specificity plays an important role in defense functions in invertebrates. In the present study, a galactose-specific lectin was detected, purified, and characterized from the hemolymph of the grub of rhinoceros beetle, Oryctes rhinoceros. The lectin was purified by affinity column chromatography with acid-treated Sepharose 6B as the matrix. The purified lectin was heat-labile, cation independent, and insensitive to EDTA. It revealed the presence of three distinct polypeptides with molecular weights of 90, 78, and 45?kDa. Thus, the native molecular weight of the purified lectin was calculated to be approximately 213?kDa, as revealed by a single band in native-PAGE. Studies on antibacterial activity and bacterial agglutination assays revealed that the purified lectin possess both these properties against two bacterial species, Bacillus subtilis and B. pumilus, isolated from the native larval habitat of O. rhinoceros.

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Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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N,N-diacetyl-glucosamine and -galactosamine derivatives as glycosyl donors

N-Acetyl-glucosamine and N-acetyl-galactosamine were converted into the O-acetyl protected 1-methylthio-derivatives 1A,B which were then transformed into N,N-diacetyl derivatives 2A and 2B, respectively. Activation of 2A,B with DMTST afforded good glycosyl donors for the generation of beta-linkages; thus, reaction with acceptors 3a,b gave oligosaccharides 4Aa and 4Ba in high and 4Ab and 4Bb in good yields. Mono-N-deacetylation could be performed with NaOMe/MeOh in quantitative yield, thus concluding a convenient procedure for the formation of beta-linked GlcNAc and GalNAc glycosides.

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Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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Distribution and molecular evolution of rhamnose-binding lectins in Salmonidae: isolation and characterization of two lectins from white-spotted Charr (Salvelinus leucomaenis) eggs.

L-Rhamnose-binding lectins were isolated from white-spotted charr (Salvelinus leucomaenis) eggs to understand the distribution and molecular evolution of the lectins in Salmonidae. Only two L-rhamnose-binding lectins, named WCL1 and WCL3, were isolated from white-spotted charr eggs, though three lectins, named STL1, STL2, and STL3, had been obtained from steelhead trout (Oncorhynchus mykiss) eggs. The cDNAs of WCL1 and WCL3 included 1,245 and 838 bp nucleotides with open reading frames of 933 and 651 nucleotides, respectively, and encoded for the complete amino acid sequences of mature proteins consisted of 288 (WCL1) and 195 (WCL3) residues, and signal sequences of 23 and 22 residues, respectively. WCLs were composed of three (for WCL1) or two (for WCL3) tandemly repeated homologous domains, which consisted of about 95 amino acid residues, and showed 91 and 93% sequence identities to STL1 and STL3, respectively. The mRNAs of WCL1 and WCL3 were detected exclusively in liver and ovary, respectively, however, neither a protein nor mRNA corresponding to STL2 could be identified in white-spotted charr. The phylogenetic tree of the sequences encoding carbohydrate recognition domains of 7 lectins from 4 species shows 5 functional clusters and their evolutional process. These results indicate that multiple L-rhamnose-binding isolectins have diverged by gene duplication and exon shuffling to play various biological roles in each species.

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Tetrahydropyran – Wikipedia,
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Spatial screening of cyclic neoglycopeptides: Identification of polyvalent wheat-germ agglutinin ligands

Affinities enhanced by up to 619-fold were found for new ligands of the plant lectin wheat-germ agglutinin (compared to the binding of N-acetylglucosamine (GlcNAc)). These polyvalent neoglycopeptides were identified from screening of a spatially diverse library of type 1. Binding affinity is determined not only by the number of GlcNAc residues but also by the ligand architecture.

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Tetrahydropyran – Wikipedia,
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Regio- and stereoselective transacylation of polyhydric alcohols using pronase in organic solvents

Pronase a mixture of proteases from Streptomyces griseus has been found to be a good catalyst for transacylation in organic solvents.With equimolar concentrations of p-nitrophenyl acetate as acyl donor in pyridine, aliphatic diols give predominantly the monoacetates.Glucose and isomeric hexoses give the 6-O-acetates in good yield. 6-Deoxy-sugars however, yield mixtures of acetates indicating that regioselectivity among the secondary hydroxyl groups are poor.Racemic trans-1,2-cyclohexanediol affords the (1R, 2R)-monoacetate in 44percent yield with greater than 90percent enantiomer ic purity. cis-1,2-cyclohexanediol and cis-1,2-cyclohexanedimethanol also give optically active monoacetates.

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Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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Natural product disaccharide engineering through tandem glycosyltransferase catalysis reversibility and neoglycosylation

A two-step strategy for disaccharide modulation using vancomycin as a model is reported. The strategy relies upon a glycosyltransferase-catalyzed ‘reverse’ reaction to enable the facile attachment of an alkoxyamine-bearing sugar to the vancomycin core. Neoglycosylation of the corresponding aglycon led to a novel set of vancomycin 1,6-disaccharide variants. While the in vitro antibacterial properties of corresponding vancomycin 1,6-disaccharide analogs were equipotent to the parent antibiotic, the chemoenzymatic method presented is expected to be broadly applicable.

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Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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Solid skin care composition comprising multiple layers

The present invention relates to a solid skin care composition comprising: (a) a first layer which is solid at 45 C. and which is a water-in-oil emulsion; and (b) a second layer which is solid at 45 C. and which is an oil-in-water emulsion comprising a benefit agent; wherein the first layer and the second layer have a different composition; and wherein the first layer and the second layer are provided in the same package in a manner such that the first layer and the second layer can be simultaneously applied.

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Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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A kind of N – acetyl glucoside compound (by machine translation)

The invention discloses a N – acetyl glucoside compound, the glucoside compound of formula (I) indicated by the structure. The glucoside compound to N – acetyl – D – glucosamine and alkoxy alcohol as raw materials, through a three-step reaction to obtain the. The invention provides compounds of formula (I) the structure shown as the glucoside compound improves the alkyl – N – acetyl glucoside water-soluble, and can greatly reduce the surface tension of the solution, has good foaming ability, foam stability and emulsifying ability, it is a cheap and easily obtained N – acetyl – D – glucosamine as a synthetic raw material obtained by chemical method with novel structure and application value of the hydrophilic surfactant. The glycoside also is expected to be in the production of hyaluronic acid, the growth of the Bifidobacterium, inhibit the propagation of Helicobacter pylori in the gastrointestinal tract of harmful role to play. In addition, in view of its novel structure, can be applied to the follow-up of the relevant product research and development and production. (by machine translation)

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Reference£º
Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics