RESUMO
Generating the barriers that protect our inner surfaces from bacteria and other challenges requires large glycoproteins called mucins. These come in two types, gel-forming and transmembrane, all characterized by large, highly O-glycosylated mucin domains that are diversely decorated by Golgi glycosyltransferases to become extended rodlike structures. The general functions of mucins on internal epithelial surfaces are to wash away microorganisms and, even more importantly, to build protective barriers. The latter function is most evident in the large intestine, where the inner mucus layer separates the numerous commensal bacteria from the epithelial cells. The host's conversion of MUC2 to the outer mucus layer allows bacteria to degrade the mucin glycans and recover the energy content that is then shared with the host. The molecular nature of the mucins is complex, and how they construct the extracellular complex glycocalyx and mucus is poorly understood and a future biochemical challenge.
Assuntos
Microbioma Gastrointestinal/fisiologia , Glicocálix/química , Glicosiltransferases/química , Células Caliciformes/química , Mucinas/química , Muco/química , Animais , Configuração de Carboidratos , Sequência de Carboidratos , Expressão Gênica , Glicocálix/metabolismo , Glicosilação , Glicosiltransferases/classificação , Glicosiltransferases/genética , Glicosiltransferases/metabolismo , Células Caliciformes/metabolismo , Células Caliciformes/microbiologia , Humanos , Mucinas/classificação , Mucinas/genética , Mucinas/metabolismo , Muco/metabolismo , Muco/microbiologia , Simbiose/fisiologiaRESUMO
Glycoscience research has been significantly impeded by the complex compositions of the glycans present in biological molecules and the lack of convenient tools suitable for studying the glycosylation process and its function. Polysaccharides and glycoconjugates are not encoded directly by genes; instead, their biosynthesis relies on the differential expression of carbohydrate enzymes, resulting in heterogeneous mixtures of glycoforms, each with a distinct physiological activity. Access to well-defined structures is required for functional study, and this has been provided by chemical and enzymatic synthesis and by the engineering of glycosylation pathways. This review covers general methods for preparing glycans commonly found in mammalian systems and applying them to the synthesis of therapeutically significant glycoconjugates (glycosaminoglycans, glycoproteins, glycolipids, glycosylphosphatidylinositol-anchored proteins) and the development of carbohydrate-based vaccines.
Assuntos
Glicoconjugados/síntese química , Glicoproteínas/síntese química , Glicosaminoglicanos/síntese química , Infecções por Haemophilus/prevenção & controle , Vacinas Anti-Haemophilus/administração & dosagem , Polissacarídeos/síntese química , Sequência de Aminoácidos , Configuração de Carboidratos , Sequência de Carboidratos , Glicoconjugados/imunologia , Glicolipídeos/síntese química , Glicolipídeos/imunologia , Glicoproteínas/imunologia , Glicosaminoglicanos/imunologia , Glicosilação , Glicosilfosfatidilinositóis/síntese química , Glicosilfosfatidilinositóis/imunologia , Infecções por Haemophilus/imunologia , Infecções por Haemophilus/microbiologia , Vacinas Anti-Haemophilus/síntese química , Haemophilus influenzae tipo b/efeitos dos fármacos , Haemophilus influenzae tipo b/crescimento & desenvolvimento , Haemophilus influenzae tipo b/patogenicidade , Humanos , Polissacarídeos/imunologiaRESUMO
A key effector route of the Sugar Code involves lectins that exert crucial regulatory controls by targeting distinct cellular glycans. We demonstrate that a single amino-acid substitution in a banana lectin, replacing histidine 84 with a threonine, significantly reduces its mitogenicity, while preserving its broad-spectrum antiviral potency. X-ray crystallography, NMR spectroscopy, and glycocluster assays reveal that loss of mitogenicity is strongly correlated with loss of pi-pi stacking between aromatic amino acids H84 and Y83, which removes a wall separating two carbohydrate binding sites, thus diminishing multivalent interactions. On the other hand, monovalent interactions and antiviral activity are preserved by retaining other wild-type conformational features and possibly through unique contacts involving the T84 side chain. Through such fine-tuning, target selection and downstream effects of a lectin can be modulated so as to knock down one activity, while preserving another, thus providing tools for therapeutics and for understanding the Sugar Code.
Assuntos
Lectinas de Plantas/química , Lectinas de Plantas/genética , Fármacos Anti-HIV/química , Sequência de Carboidratos , Engenharia Genética , Mitógenos/química , Modelos Moleculares , Simulação de Dinâmica Molecular , Musa/químicaRESUMO
In the last decade, glycan microarrays have revolutionized the analysis of the specificity of glycan-binding proteins (GBPs), providing information that simultaneously illuminates the biology mediated by them and decodes the informational content of the glycome. Numerous methods have emerged for arraying glycans in a "chip" format, and glycan libraries have been assembled that address the diversity of the human glycome. Such arrays have been successfully used for analysis of GBPs, which mediate mammalian biology, host-pathogen interactions, and immune recognition of glycans relevant to vaccine production and cancer antigens. This review covers the development of glycan microarrays and applications that have provided insights into the roles of mammalian and microbial GBPs.
Assuntos
Glicômica/métodos , Análise em Microsséries/métodos , Polissacarídeos/análise , Animais , Configuração de Carboidratos , Sequência de Carboidratos , Humanos , Dados de Sequência Molecular , Estrutura Molecular , Lectinas de Plantas/química , Lectinas de Plantas/metabolismo , Ligação Proteica , Proteínas/química , Proteínas/metabolismoRESUMO
The peptidoglycan cell wall is a macromolecular structure that encases bacteria and is essential for their survival. Proper assembly of the cell wall requires peptidoglycan synthases as well as membrane-bound cleavage enzymes that control where new peptidoglycan is made and inserted. Previous studies have shown that two membrane-bound proteins in Streptococcus pneumoniae, here named MpgA and MpgB, are important in maintaining cell wall integrity. MpgA was predicted to be a lytic transglycosylase based on its homology to Escherichia coli MltG, while the enzymatic activity of MpgB was unclear. Using nascent peptidoglycan substrates synthesized in vitro from the peptidoglycan precursor Lipid II, we report that both MpgA and MpgB are muramidases. We show that replacing a single amino acid in E. coli MltG with the corresponding amino acid from MpgA results in muramidase activity, allowing us to predict from the presence of this amino acid that other putative lytic transglycosylases actually function as muramidases. Strikingly, we report that MpgA and MpgB cut nascent peptidoglycan at different positions along the sugar backbone relative to the reducing end, with MpgA producing much longer peptidoglycan oligomers. We show that the cleavage site selectivity of MpgA is controlled by the LysM-like subdomain, which is required for its full functionality in cells. We propose that MltG's ability to complement the loss of MpgA in S. pneumoniae despite performing different cleavage chemistry is because it can cleave nascent peptidoglycan at the same distance from the lipid anchor.
Assuntos
Proteínas de Bactérias/metabolismo , Parede Celular/enzimologia , Glicosídeo Hidrolases/metabolismo , Streptococcus pneumoniae/metabolismo , Substituição de Aminoácidos , Sequência de Carboidratos , Hidrólise , Peptidoglicano/química , Peptidoglicano/metabolismoRESUMO
The application of solid-state (SS) nanopore devices to single-molecule nucleic acid sequencing has been challenging. Thus, the early successes in applying SS nanopore devices to the more difficult class of biopolymer, glycosaminoglycans (GAGs), have been surprising, motivating us to examine the potential use of an SS nanopore to analyze synthetic heparan sulfate GAG chains of controlled composition and sequence prepared through a promising, recently developed chemoenzymatic route. A minimal representation of the nanopore data, using only signal magnitude and duration, revealed, by eye and image recognition algorithms, clear differences between the signals generated by four synthetic GAGs. By subsequent machine learning, it was possible to determine disaccharide and even monosaccharide composition of these four synthetic GAGs using as few as 500 events, corresponding to a zeptomole of sample. These data suggest that ultrasensitive GAG analysis may be possible using SS nanopore detection and well-characterized molecular training sets.
Assuntos
Heparitina Sulfato/química , Aprendizado de Máquina , Nanoporos , Sequência de Carboidratos , Dissacarídeos/química , Glicômica/métodos , Glicômica/normas , Heparitina Sulfato/síntese química , Monossacarídeos/químicaRESUMO
Humans express seven heparan sulfate (HS) 3-O-sulfotransferases that differ in substrate specificity and tissue expression. Although genetic studies have indicated that 3-O-sulfated HS modulates many biological processes, ligand requirements for proteins engaging with HS modified by 3-O-sulfate (3-OS) have been difficult to determine. In particular, the context in which the 3-OS group needs to be presented for binding is largely unknown. We describe herein a modular synthetic approach that can provide structurally diverse HS oligosaccharides with and without 3-OS. The methodology was employed to prepare 27 hexasaccharides that were printed as a glycan microarray to examine ligand requirements of a wide range of HS-binding proteins. The binding selectivity of antithrombin-III (AT-III) compared well with anti-Factor Xa activity supporting robustness of the array technology. Many of the other examined HS-binding proteins required an IdoA2S-GlcNS3S6S sequon for binding but exhibited variable dependence for the 2-OS and 6-OS moieties, and a GlcA or IdoA2S residue neighboring the central GlcNS3S. The HS oligosaccharides were also examined as inhibitors of cell entry by herpes simplex virus type 1, which, surprisingly, showed a lack of dependence of 3-OS, indicating that, instead of glycoprotein D (gD), they competitively bind to gB and gC. The compounds were also used to examine substrate specificities of heparin lyases, which are enzymes used for depolymerization of HS/heparin for sequence determination and production of therapeutic heparins. It was found that cleavage by lyase II is influenced by 3-OS, while digestion by lyase I is only affected by 2-OS. Lyase III exhibited sensitivity to both 3-OS and 2-OS.
Assuntos
Células Epiteliais/metabolismo , Heparina Liase/metabolismo , Heparitina Sulfato/metabolismo , Herpesvirus Humano 1/metabolismo , Sulfatos/metabolismo , Sulfotransferases/metabolismo , Acetilglucosamina/química , Acetilglucosamina/metabolismo , Antitrombina III/química , Antitrombina III/genética , Antitrombina III/metabolismo , Sítios de Ligação , Ligação Competitiva , Sequência de Carboidratos , Linhagem Celular , Córnea/citologia , Córnea/metabolismo , Células Epiteliais/patologia , Células Epiteliais/virologia , Fator Xa/química , Fator Xa/genética , Fator Xa/metabolismo , Inibidores do Fator Xa/química , Inibidores do Fator Xa/metabolismo , Expressão Gênica , Ácido Glucurônico/química , Ácido Glucurônico/metabolismo , Heparina Liase/química , Heparina Liase/genética , Heparitina Sulfato/química , Herpesvirus Humano 1/crescimento & desenvolvimento , Interações Hospedeiro-Patógeno/genética , Humanos , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/metabolismo , Análise em Microsséries , Ligação Proteica , Proteólise , Bibliotecas de Moléculas Pequenas , Especificidade por Substrato , Sulfatos/química , Sulfotransferases/química , Sulfotransferases/genética , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/metabolismoRESUMO
All members of the family Chlamydiaceae have lipopolysaccharides (LPS) that possess a shared carbohydrate trisaccharide antigen, 3-deoxy-d-manno-oct-2-ulosonic acid (Kdo) that is functionally uncharacterized. A single gene, genus-specific epitope (gseA), is responsible for attaching the tri-Kdo to lipid IVA. To investigate the function of Kdo in chlamydial host cell interactions, we made a gseA-null strain (L2ΔgseA) by using TargeTron mutagenesis. Immunofluorescence microscopy and immunoblotting with a Kdo-specific monoclonal antibody demonstrated that L2ΔgseA lacked Kdo. L2ΔgseA reacted by immunoblotting with a monoclonal antibody specific for a conserved LPS glucosamine-PO4 epitope, indicating that core lipid A was retained by the mutant. The mutant strain produced a similar number of inclusions as the parental strain but yielded lower numbers of infectious elementary bodies. Transmission electron microscopy of L2ΔgseA-infected cells showed atypical developmental forms and a reduction in the number of elementary bodies. Immunoblotting of dithiothreitol-treated L2ΔgseA-infected cells lysates revealed a marked reduction in outer membrane OmcB disulfide cross-linking, suggesting that the elementary body outer membrane structure was affected by the lack of Kdo. Notably, lactic acid dehydrogenase release by infected cells demonstrated that L2ΔgseA was significantly more cytotoxic to host cells than the wild type. The cytotoxic phenotype may result from an altered outer membrane biogenesis structure and/or function or, conversely, from a direct pathobiological effect of Kdo on an unknown host cell target. These findings implicate a previously unrecognized role for Kdo in host cell interactions that facilitates postinfection host cell survival.
Assuntos
Chlamydia trachomatis , Lipopolissacarídeos , Lipopolissacarídeos/metabolismo , Sequência de Carboidratos , Epitopos , Açúcares Ácidos , Anticorpos MonoclonaisRESUMO
The synthesis of defined oligosaccharides is a complex task. Several enabling technologies have been introduced in the last two decades to facilitate synthetic access to these valuable biomolecules. In this concept, we describe the technological solutions that have advanced glycochemistry using automated glycan assembly, flow chemistry and data science as examples. We highlight how the synergies between these different technologies can further advance the field, with progress toward the realization of a self-driving lab for glycan synthesis.
Assuntos
Ciência de Dados , Polissacarídeos , Glicosilação , Sequência de Carboidratos , Polissacarídeos/química , Oligossacarídeos/químicaRESUMO
Mycobacteria are covered in a thick layer of different polysaccharides that helps to avert the innate immune response. Lipoarabinomannan (LAM) and arabinogalactan (AG) are ubiquitously contained in these envelopes, and rapid access to defined oligo- and polysaccharides is essential to elucidate their structural and biological roles. Arabinofuranose (Araf) residues in LAM and AG are connected either via α-1,2-trans linkages that are synthetically straightforward to install or the more challenging ß-(1,2-cis) linkages. Herein, it was demonstrated that automated glycan assembly (AGA) can be used to quickly prepare 1,2-cis-ß-Araf as illustrated by the assembly of a highly branched arabinan hexasaccharide and a docosasaccharide arabinan (Araf23 ) motif.
Assuntos
Mycobacterium tuberculosis , Mycobacterium tuberculosis/química , Sequência de Carboidratos , Polissacarídeos/química , Lipopolissacarídeos/químicaRESUMO
The synthesis of a disaccharide macrocycle through 2,3-dideoxy glucopyranosyl monosaccharide is reported. 2,3-Dideoxy-erythro-hexopyranosyl thioglycoside possessing a free hydroxy functionality at the C-4 carbon is prepared, and cycloglycosylation is conducted. In the event, the cycloglycosylation occurs with a ring contraction of the monosaccharide moiety and affords the cyclic furanoside disaccharide. Solution-phase and single-crystal X-ray diffraction structural characterizations permit the features of the macrocycle to be uncovered. The solubilization and encapsulation properties of the macrocycle are studied in aqueous solutions with 1-aminoadamantane.
Assuntos
Dissacarídeos , Glicosídeos , Glicosídeos/química , Sequência de Carboidratos , Dissacarídeos/química , Cristalografia por Raios X , MonossacarídeosRESUMO
Herein, we report a concise synthetic approach for the first total synthesis of a pentasaccharide repeating unit of Acinetobacter baumannii K11 capsular polysaccharides containing a rare sugar 6-deoxy-l-talose. The pentasaccharide was synthesized in a convergent manner using a [3 + 2] block glycosylation strategy. During this synthetic strive, we used a 2,2,2-trichloroethoxycarbonyl (Troc)-protected monosaccharide unit to achieve a high yield during the glycosylation to synthesize a trisaccharide, and chemoselective deprotection of the Troc group from the trisaccharide was carried out under a mild, pH-neutral condition, keeping the O-glycosidic bond, azido, and acid/base sensitive group intact. A thiotolylglycoside disaccharide donor containing 6-deoxy-l-talose was synthesized for the first time by the armed-disarmed glycosylation method between two thiotolylglycosides.
Assuntos
Acinetobacter baumannii , Acinetobacter baumannii/química , Sequência de Carboidratos , Polissacarídeos/química , Oligossacarídeos/química , Trissacarídeos/química , Polissacarídeos Bacterianos/químicaRESUMO
Chondroitin sulfate (CS), a linear acidic polysaccharide, exhibits numerous biological activities that are dependent on sulfation patterns. CS oligosaccharides comprise repeating disaccharide units with different (hetero)-type sulfation patterns and are common in nature. We herein report the synthesis of the following biotinylated CS tetrasaccharides: CS-AD [ßGalNAc4S(1-4)ßGlcA(1-3)ßGalNAc6S(1-4)ßGlcA2S] and CS-DA [ßGalNAc6S(1-4)ßGlcA2S(1-3)ßGalNAc4S(1-4)ßGlcA], in a stereo-controlled manner. We also demonstrated that the CS-d-specific monoclonal antibody MO-225 bound more strongly to CS-DA than to CS-DD or -AD.
Assuntos
Sulfatos de Condroitina , Dissacarídeos , Sequência de Carboidratos , Oligossacarídeos , Anticorpos MonoclonaisRESUMO
Synthesis of glycosaminoglycans, such as heparan sulfate (HS) and chondroitin sulfate (CS), occurs in the lumen of the Golgi, but the relationship between Golgi structural integrity and glycosaminoglycan synthesis is not clear. In this study, we disrupted the Golgi structure by knocking out GRASP55 and GRASP65 and determined its effect on the synthesis, sulfation, and secretion of HS and CS. We found that GRASP depletion increased HS synthesis while decreasing CS synthesis in cells, altered HS and CS sulfation, and reduced both HS and CS secretion. Using proteomics, RNA-seq and biochemical approaches, we identified EXTL3, a key enzyme in the HS synthesis pathway, whose level is upregulated in GRASP knockout cells; while GalNAcT1, an essential CS synthesis enzyme, is robustly reduced. In addition, we found that GRASP depletion decreased HS sulfation via the reduction of PAPSS2, a bifunctional enzyme in HS sulfation. Our study provides the first evidence that Golgi structural defect may significantly alter the synthesis and secretion of glycosaminoglycans.
Assuntos
Glicosaminoglicanos/metabolismo , Complexo de Golgi/metabolismo , Proteínas da Matriz do Complexo de Golgi/fisiologia , Metabolismo dos Carboidratos/genética , Sequência de Carboidratos/genética , Sulfatos de Condroitina/química , Sulfatos de Condroitina/metabolismo , Deleção de Genes , Técnicas de Silenciamento de Genes , Complexo de Golgi/patologia , Proteínas da Matriz do Complexo de Golgi/genética , Células HeLa , Heparitina Sulfato/química , Heparitina Sulfato/metabolismo , Humanos , Via Secretória/genética , Sulfatos/metabolismoRESUMO
ß1-3/1-6 Glucans, known for their diverse structures, comprise a ß1-3-linked main chain and ß1-6-linked short branches. Laminarin, a ß1-3/1-6 glucan extracted from brown seaweed, for instance, includes ß1-6 linkages even in the main chain. The diverse structures provide various beneficial functions for the glucan. To investigate the relationship between structure and functionality, and to enable the characterization of ß1-3/1-6 glucan-metabolizing enzymes, oligosaccharides containing the exact structures of ß1-3/1-6 glucans are required. We synthesized the monomeric units for the synthesis of ß1-3/1-6 mixed-linked glucooligosaccharides. 2-(Trimethylsilyl)ethyl 2-O-benzoyl-4,6-O-benzylidene-ß-d-glucopyranoside served as an acceptor in the formation of ß1-3 linkages. Phenyl 2-O-benzoyl-4,6-O-benzylidene-3-O-(tert-butyldiphenylsilyl)-1-thio-ß-d-glucopyranoside and phenyl 2,3-di-O-benzoyl-4,6-di-O-levulinyl-1-thio-ß-d-glucopyranoside acted as donors, synthesizing acceptors suitable for the formation of ß1-3- and ß1-6-linkages, respectively. These were used to synthesize a derivative of Glcß1-6Glcß1-3Glcß1-3Glc, demonstrating that the proposed route can be applied to synthesize the main chain of ß-glucan, with the inclusion of both ß1-3 and ß1-6 linkages.
Assuntos
Glucosídeos , beta-Glucanas , Sequência de Carboidratos , Oligossacarídeos/química , GlucanosRESUMO
A novel acidic heteropolysaccharide (LCP-90-1) was isolated and purified from a traditional "heat-clearing" Chinese medicine, Lysimachia christinae Hance. LCP-90-1 (Mw, 20.65 kDa) was composed of Man, Rha, GlcA, Glc, Gal, and Ara, with relative molar ratios of 1.00: 3.00: 11.62: 1.31: 1.64: 5.24. The backbone consisted of 1,4-α-D-GlcpA, 1,4-α-D-Glcp, 1,4-ß-L-Rhap, and 1,3,5-α-L-Araf, with three branches of ß-D-Galp-(1 â 4)-ß-L-Rhap-(1â, α-L-Araf-(1â and α-D-Manp-(1â attached to the C-5 position of 1,3,5-α-L-Araf. LCP-90-1 exhibited potent anticomplement activity (CH50: 135.01 ± 0.68 µg/mL) in vitro, which was significantly enhanced with increased glucuronic acid (GlcA) content in its degradation production (LCP-90-1-A, CH50: 28.26 ± 0.39 µg/mL). However, both LCP-90-1 and LCP90-1-A were inactivated after reduction or complete acid hydrolysis. These observations indicated the important role of GlcA in LCP-90-1 and associated derivatives with respect to anticomplement activity. Similarly, compared with LCP-90-1, the antioxidant activity of LCP-90-1-A was also enhanced. Thus, polysaccharides with a high content of GlcA might be important and effective substances of L. christinae.
Assuntos
Lysimachia , Polissacarídeos , Humanos , Sequência de Carboidratos , Polissacarídeos/química , Hidrólise , Ácido GlucurônicoRESUMO
Several sialoglycopeptides were isolated from several fish eggs and exerted anti-osteoporosis effects. However, few papers have explored sialoglycopeptide from tuna eggs (T-ES). Here, a novel T-ES was prepared through extraction with KCl solution and subsequent enzymolysis. Pure T-ES was obtained through DEAE-Sepharose ion exchange chromatography and sephacryl S-300 gel filtration chromatography. The T-ES was composed of 14.07% protein, 73.54% hexose, and 8.28% Neu5Ac, with a molecular weight of 9481 Da. The backbone carbohydrate in the T-ES was â4)-ß-D-GlcN-(1â3)-α-D-GalN-(1â3)-ß-D-Glc-(1â2)-α-D-Gal-(1â2)-α-D-Gal-(1â3)-α-D-Man-(1â, with two branches of ß-D-GlcN-(1â and α-D-GalN-(1â linking at o-4 in â2,4)-α-D-Gal-(1â. Neu5Ac in the T-ES was linked to the branch of α-D-GlcN-(1â. A peptide chain, Ala-Asp-Asn-Lys-Ser*-Met-Ile that was connected to the carbohydrate chain through O-glycosylation at the -OH of serine. Furthermore, in vitro data revealed that T-ES could remarkably enhance bone density, bone biomechanical properties, and bone microstructure in SAMP mice. The T-ES elevated serum osteogenesis-related markers and reduced bone resorption-related markers in serum and urine. The present study's results demonstrated that T-ES, a novel sialoglycopeptide, showed significant anti-osteoporosis effects, which will accelerate the utilization of T-ES as an alternative marine drug or functional food for anti-osteoporosis.
Assuntos
Sialoglicoproteínas , Atum , Humanos , Camundongos , Animais , Sequência de Carboidratos , Carboidratos , HexosesRESUMO
Two closely related Proteus mirabilis smooth strains, Kr1 and Ks20, were isolated from wound and skin samples, respectively, of two infected patients in central Poland. Serological tests, using the rabbit Kr1-specific antiserum, revealed that both strains presented the same O serotype. Their O antigens are unique among the Proteus O serotypes, which had been described earlier, as they were not recognized in an enzyme-linked immunosorbent assay (ELISA) by a set of Proteus O1-O83 antisera. Additionally, the Kr1 antiserum did not react with O1-O83 lipopolysaccharides (LPSs). The O-specific polysaccharide (OPS, O antigen) of P. mirabilis Kr1 was obtained via the mild acid degradation of the LPSs, and its structure was established via a chemical analysis and one- and two-dimensional 1H and 13C nuclear magnetic resonance (NMR) spectroscopy applied to both initial and O-deacetylated polysaccharides, where most ß-2-acetamido-2-deoxyglucose (N-acetylglucosamine) (GlcNAc) residues are non-stoichiometrically O-acetylated at positions 3, 4, and 6 or 3 and 6, and a minority of α-GlcNAc residues are 6-O-acetylated. Based on the serological features and chemical data, P. mirabilis Kr1 and Ks20 were proposed as candidates to a new successive O-serogroup in the genus Proteus, O84, which is another example of new Proteus O serotypes identified lately among serologically differentiated Proteus bacilli infecting patients in central Poland.
Assuntos
Antígenos O , Proteus mirabilis , Animais , Coelhos , Antígenos O/química , Sorogrupo , Sequência de Carboidratos , Proteus , Lipopolissacarídeos , SorotipagemRESUMO
The Edwardsiella genus presents five different pathogenic species: Edwardsiella tarda, E. anguillarum, E. piscicida, E. hoshinae and E. ictaluri. These species cause infections mainly in fish, but they can also infect reptiles, birds or humans. Lipopolysaccharide (endotoxin) plays an important role in the pathogenesis of these bacteria. For the first time, the chemical structure and genomics of the lipopolysaccharide (LPS) core oligosaccharides of E. piscicida, E. anguillarum, E. hoshinae and E. ictaluri were studied. The complete gene assignments for all core biosynthesis gene functions were acquired. The structure of core oligosaccharides was investigated by ¹H and 13C nuclear magnetic resonance (NMR) spectroscopy. The structures of E. piscicida and E. anguillarum core oligosaccharides show the presence of â3,4)-L-glycero-α-D-manno-Hepp, two terminal ß-D-Glcp, â2,3,7)-L-glycero-α-D-manno-Hepp, â7)-L-glycero-α-D-manno-Hepp, terminal α-D-GlcpN, two â4)-α-D-GalpA, â 3)-α-D-GlcpNAc, terminal ß-D-Galp and â5-substituted Kdo. E. hoshinare core oligosaccharide shows only one terminal ß-D-Glcp, and instead of terminal ß-D-Galp a terminal α-D-GlcpNAc. E. ictaluri core oligosaccharide shows only one terminal ß-D-Glcp, one â4)-α-D-GalpA and do not have terminal α-D-GlcpN (see complementary figure).
Assuntos
Lipopolissacarídeos , Oligossacarídeos , Animais , Humanos , Sequência de Carboidratos , Lipopolissacarídeos/química , Oligossacarídeos/química , Espectroscopia de Ressonância Magnética , MetilaçãoRESUMO
The system of the four different human blood groups is based on the oligosaccharide antigens A or B, which are located on the surface of blood cells and other cells including endothelial cells, attached to the membrane proteins or lipids. After transfusion, the presence of these antigens on the apical surface of endothelial cells could induce an immunological reaction against the host. The final oligosaccharide sequence of AgA consists of Gal-GlcNAc-Gal (GalNAc)-Fuc. AgB contains Gal-GlcNAc-Gal (Gal)-Fuc. These antigens are synthesised in the Golgi complex (GC) using unique Golgi glycosylation enzymes (GGEs). People with AgA also synthesise antibodies against AgB (group A [II]). People with AgB synthesise antibodies against AgA (group B [III]). People expressing AgA together with AgB (group AB [IV]) do not have these antibodies, while people who do not express these antigens (group O [0; I]) synthesise antibodies against both antigens. Consequently, the antibodies are synthesised against antigens that apparently do not exist in the body. Here, we compared the prediction power of the main hypotheses explaining the formation of these antibodies, namely, the concept of natural antibodies, the gut bacteria-derived antibody hypothesis, and the antibodies formed as a result of glycosylation mistakes or de-sialylation of polysaccharide chains. We assume that when the GC is overloaded with lipids, other less specialised GGEs could make mistakes and synthesise the antigens of these blood groups. Alternatively, under these conditions, the chylomicrons formed in the enterocytes may, under this overload, linger in the post-Golgi compartment, which is temporarily connected to the endosomes. These compartments contain neuraminidases that can cleave off sialic acid, unmasking these blood antigens located below the acid and inducing the production of antibodies.