The anomeric configurations of the two ammonium (methyl 3-deoxy-d-manno-2-octulopyranosid)onate salts (methyl α- and β-ketopyranosides of KDO)

Unger, Frank M.; Stix, Diana; Schulz, Gerhard

doi:10.1016/s0008-6215(00)85325-2

Cited by 109 publications

(48 citation statements)

References 8 publications

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“…The complete assignment of 1 H and 13 C NMR signals of 3 was performed using a combination of 1 H NMR, proton-coupled 13 C NMR, two-dimensional 1 H, 1 H correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), rotating-frame Overhauser enhancement spectroscopy (ROESY), 1 H, 13 C heteronuclear single quantum coherence (HSQC), and heteronuclear multiple bond correlation (HMBC) experiments. The HSQC spectrum of product 3 ( Correlations from H-3 to H-5 were traced in the COSY and TOCSY spectra, and the signals for corresponding carbons were assigned by HSQC.…”

Section: N-terminal Domain Of Kpsc Catalyzes Addition Of a Single ␤-(mentioning

confidence: 99%

“…The 13 C NMR chemical shifts of residue B were indicative of its terminal position, whereas residue A C-4 signal at ␦ 73.3 was shifted downfield compared with its position at ␦ 68.7 in acceptor 2, because of the ␣-glycosylation effect. Based on 13 C NMR data and the correlation with 1 H signals in HMBC spectrum, i.e. C-2/O-CH 2 -C at ␦ 102.3/3.39 and 102.3/3.66 ( Fig.…”

Section: N-terminal Domain Of Kpsc Catalyzes Addition Of a Single ␤-(mentioning

confidence: 99%

“…The signals for residue A were close to those for the 34)-substituted Kdo residue A in product 3, whereas the signals for residue C are indicative of its terminal position. The substitution of residue B in 4 at position O-7 was inferred from a positive ␣-glycosylation effect on C-7 and the negative ␤-glycosylation effect on neighboring carbons in residue B (C-6 and C-8) compared with 13 C NMR signals of the terminal Kdo residue in 3. The HMBC spectrum showed the correlation between residue C C-2 and residue B H-7 at ␦ 101.9/4.40, which, taking in consideration the substitution positions, determined the sequence of Kdo residue as follows: C(237)B(234)A.…”

Section: C-terminal Domain Of Kpsc Catalyzes Addition Of a Single ␤-(mentioning

confidence: 99%

“…NMR structural analysis of in vitro enzymatic products 3 (left panels) and 4 (right panels). Top panels, selected regions of 1 H, 13 C HSQC spectra, showing correlations from carbohydrate moieties. Numbers refer to H/C pairs in the Kdo residues designated by letters.…”

Section: ␤-Kdo Oligosaccharide Consists Of Alternating ␤-(234)-and ␤-mentioning

confidence: 99%

“…The asterisk indicates the signal from a PEG impurity. Bottom panels, selected regions of the 1 H, 13 C HMBC spectra, demonstrating intra-and inter-residue correlations from the Kdo anomeric carbons (C-2). KpsC ␤-Kdo Glycosyltransferase Activity OCTOBER 7, 2016 • VOLUME 291 • NUMBER 41 the structure of Kdo linker, the products 3 and 4 were used as acceptors in reactions with purified KpsC and a 20-fold molar excess of donor substrate.…”

Section: ␤-Kdo Oligosaccharide Consists Of Alternating ␤-(234)-and ␤-mentioning

confidence: 99%

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Biochemical Characterization of Bifunctional 3-Deoxy-β-d-manno-oct-2-ulosonic Acid (β-Kdo) Transferase KpsC from Escherichia coli Involved in Capsule Biosynthesis

Ovchinnikova

Doyle

Huang

et al. 2016

Journal of Biological Chemistry

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3-Deoxy-D-manno-oct-2-ulosonic acid (Kdo) is an essential component of bacterial lipopolysaccharides, where it provides the linkage between lipid and carbohydrate moieties. In all known LPS structures, Kdo residues possess ␣-anomeric configurations, and the corresponding inverting ␣-Kdo transferases are well characterized. Recently, it has been shown that a large group of capsular polysaccharides from Gram-negative bacteria, produced by ATP-binding cassette transporter-dependent pathways, are also attached to a lipid anchor through a conserved Kdo oligosaccharide. In the study reported here, the structure of this Kdo linker was determined by NMR spectroscopy, revealing alternating ␤-(234)-and ␤-(237)-linked Kdo residues. KpsC contains two retaining ␤-Kdo glycosyltransferase domains belonging to family GT99 that are responsible for polymerizing the ␤-Kdo linker on its glycolipid acceptor. Full-length Escherichia coli KpsC was expressed and purified, together with the isolated N-terminal domain and a mutant protein (KpsC D160A) containing a catalytically inactivated N-terminal domain. The Kdo transferase activities of these proteins were determined in vitro using synthetic acceptors, and the reaction products were characterized using TLC, mass spectrometry, and NMR spectroscopy. The N-and C-terminal domains were found to catalyze formation of ␤-(234) and ␤-(237) linkages, respectively. Based on phylogenetic analyses, we propose the linkage specificities of the glycosyltransferase domains are conserved in KpsC homologs from other bacterial species.Pathogenic bacteria are often covered by a protective layer of polysaccharide known as the capsule. Capsules are important virulence determinants because, depending on the pathogen, they can protect bacteria against phagocytosis and complement-mediated killing, as well as helping in adherence to host cells (1). In Gram-negative bacteria, capsular polysaccharides (CPS) 4 are synthesized and exported onto the surface of the outer membrane via one of two widely disseminated and fundamentally different assembly systems (1, 2). The ATP-binding cassette (ABC) transporter-dependent system is the focus of this study. In the nomenclature of Escherichia coli, these systems generate "group 2" capsules (1, 2). In group 2 CPS assembly, biosynthesis is initiated by KpsS, which transfers a single 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) residue from its precursor (CMP-␤-Kdo) to (lyso)phosphatidylglycerol, in a reaction located at the cytoplasm-membrane interface (1). The KpsS product is then extended by KpsC to form the ␤-Kdo linker, which typically consists of five to nine Kdo residues (3). The serotype-specific glycosyltransferases (GTs) extend this linker to complete polymerization of CPS in the cytoplasm before it is exported by the pathway-defining ABC transporter and then translocated to the cell surface (1, 2).The native lipid-linked ␤-Kdo-oligosaccharides were isolated from E. coli K1 and K5 and Neisseria meningitidis group B by enzymatic depolymerization of high molecular ma...

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Section: N-terminal Domain Of Kpsc Catalyzes Addition Of a Single ␤-(mentioning

confidence: 99%

Section: N-terminal Domain Of Kpsc Catalyzes Addition Of a Single ␤-(mentioning

confidence: 99%

Section: C-terminal Domain Of Kpsc Catalyzes Addition Of a Single ␤-(mentioning

confidence: 99%

Section: ␤-Kdo Oligosaccharide Consists Of Alternating ␤-(234)-and ␤-mentioning

confidence: 99%

Section: ␤-Kdo Oligosaccharide Consists Of Alternating ␤-(234)-and ␤-mentioning

confidence: 99%

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Biochemical Characterization of Bifunctional 3-Deoxy-β-d-manno-oct-2-ulosonic Acid (β-Kdo) Transferase KpsC from Escherichia coli Involved in Capsule Biosynthesis

Ovchinnikova

Doyle

Huang

et al. 2016

Journal of Biological Chemistry

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Einfache Synthese von KDO‐α‐Glycosiden durch anomer selektive O‐Alkylierung

Schmidt

Esswein

1988

Angewandte Chemie

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Arbeitsvorschriftvilllig andere Eigenschaften auf als 4 (NMR. TLC); P. Mosset, R. Gree, 4 : Zu einer auf -80°C gekuhlten Lasung von 361 mg (1.29 mmol) (-)-Z in 10 mL wasserfreiem Dichlormethan gibt man 0.72 mL (5 Aquiv.) TiCL und 15 min splter zu der nunmehr roten LBsung 470 mg (2.6 mmol) des Allens 3. Man riihrt 130 min bei -70°C und gieOt dann die dunkelrote Usung in eine kriftig gerilhrte Mischung von 50 m L Ether und 50 mL 1 N wi7Origer Bicar-Q VCH Verlagsgesellschaft mbH. D-6940 Weinheim. 1988 0044-8249/88/0909-1234 $ 02.50/0 Angew. Chem. 100 (1988) Nr. 9

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Stereoselective Synthesis of Oligo‐α(2,8)‐3‐deoxy‐D‐manno‐2‐octulosonic Acid Derivatives

Tanaka¹,

Takahashi²,

Takahashi³

2006

Angewandte Chemie

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The anomeric configurations of the two ammonium (methyl 3-deoxy-d-manno-2-octulopyranosid)onate salts (methyl α- and β-ketopyranosides of KDO)

Cited by 109 publications

References 8 publications

Biochemical Characterization of Bifunctional 3-Deoxy-β-d-manno-oct-2-ulosonic Acid (β-Kdo) Transferase KpsC from Escherichia coli Involved in Capsule Biosynthesis

Biochemical Characterization of Bifunctional 3-Deoxy-β-d-manno-oct-2-ulosonic Acid (β-Kdo) Transferase KpsC from Escherichia coli Involved in Capsule Biosynthesis

Einfache Synthese von KDO‐α‐Glycosiden durch anomer selektive O‐Alkylierung

Stereoselective Synthesis of Oligo‐α(2,8)‐3‐deoxy‐D‐manno‐2‐octulosonic Acid Derivatives

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