Poly-N-acetylglucosamine and poly(glycerol phosphate) teichoic acid identification from staphylococcal biofilm extracts using excitation sculptured TOCSY NMR

2008 ◽  
Vol 4 (2) ◽  
pp. 170-174 ◽  
Author(s):  
Jane L. Wagstaff ◽  
Irina Sadovskaya ◽  
Evgeny Vinogradov ◽  
Saïd Jabbouri ◽  
Mark J. Howard
Keyword(s):  
Teichoic Acid ◽  
Glycerol Phosphate

scholarly journals Surface Glycopolymers Are Crucial forIn VitroAnti-Wall Teichoic Acid IgG-Mediated Complement Activation and Opsonophagocytosis of Staphylococcus aureus

2015 ◽  
Vol 83 (11) ◽  
pp. 4247-4255 ◽  
Author(s):  
Jong-Ho Lee ◽  
Na-Hyang Kim ◽  
Volker Winstel ◽  
Kenji Kurokawa ◽  
Jesper Larsen ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Teichoic Acid ◽  
Complement C3 ◽  
Glycerol Phosphate ◽  
Glycosylation Pattern ◽  
Wall Teichoic Acid ◽  
Content Type ◽  
Gram Positive Bacteria ◽  
Capsule Production ◽  
Cell Envelopes

ABSTRACTThe cell envelopes of many Gram-positive bacteria contain wall teichoic acids (WTAs).Staphylococcus aureusWTAs are composed of ribitol phosphate (RboP) or glycerol phosphate (GroP) backbones substituted withd-alanine andN-acetyl-d-glucosamine (GlcNAc) orN-acetyl-d-galactosamine (GalNAc). Two WTA glycosyltransferases, TarM and TarS, are responsible for modifying the RboP WTA with α-GlcNAc and β-GlcNAc, respectively. We recently reported that purified human serum anti-WTA IgG specifically recognizes β-GlcNAc of the staphylococcal RboP WTA and then facilitates complement C3 deposition and opsonophagocytosis ofS. aureuslaboratory strains. This prompted us to examine whether anti-WTA IgG can induce C3 deposition on a diverse set of clinicalS. aureusisolates. To this end, we compared anti-WTA IgG-mediated C3 deposition and opsonophagocytosis abilities using 13 different staphylococcal strains. Of note, the majority ofS. aureusstrains tested was recognized by anti-WTA IgG, resulting in C3 deposition and opsonophagocytosis. A minority of strains was not recognized by anti-WTA IgG, which correlated with either extensive capsule production or an alteration in the WTA glycosylation pattern. Our results demonstrate that the presence of WTAs with TarS-mediated glycosylation with β-GlcNAc in clinically isolatedS. aureusstrains is an important factor for induction of anti-WTA IgG-mediated C3 deposition and opsonophagocytosis.

scholarly journals Use of CDP-Glycerol as an Alternate Acceptor for the Teichoic Acid Polymerase Reveals that Membrane Association Regulates Polymer Length

2008 ◽  
Vol 190 (21) ◽  
pp. 6940-6947 ◽  
Author(s):  
Jeffrey W. Schertzer ◽  
Eric D. Brown
Keyword(s):  
Isotope Labeling ◽  
Teichoic Acid ◽  
Extracellular Polysaccharide ◽  
Membrane System ◽  
Polymer Synthesis ◽  
Glycerol Phosphate ◽  
Polysaccharide Biosynthesis ◽  
Steady State Kinetics ◽  
Length Regulation ◽  
Lag Period

ABSTRACT The study of bacterial extracellular polysaccharide biosynthesis is hampered by the fact that these molecules are synthesized on membrane-resident carrier lipids. To get around this problem, a practical solution has been to synthesize soluble lipid analogs and study the biosynthetic enzymes using a soluble system. This has been done for the Bacillus subtilis teichoic acid polymerase, TagF, although several aspects of catalysis were inconsistent with the results obtained with reconstituted membrane systems or physiological observations. In this work we explored the acceptor substrate promiscuity and polymer length disregulation that appear to be characteristic of TagF activity away from biological membranes. Using isotope labeling, steady-state kinetics, and chemical lability studies, we demonstrated that the enzyme can synthesize poly(glycerol phosphate) teichoic acid using the elongation substrate CDP-glycerol as an acceptor. This suggests that substrate specificity is relaxed in the region distal to the glycerol phosphate moiety in the acceptor molecule under these conditions. Polymer synthesis proceeded at a rate (27 min−1) comparable to that in the reconstituted membrane system after a distinct lag period which likely represented slower initiation on the unnatural CDP-glycerol acceptor. We confirmed that polymer length became disregulated in the soluble system as the polymers synthesized on CDP-glycerol acceptors were much larger than the polymers synthesized on the membrane or previously found attached to bacterial cell walls. Finally, polymer synthesis on protease-treated membranes suggested that proper length regulation is retained in the absence of accessory proteins and provided evidence that such regulation is conferred through proper association of the polymerase with the membrane.

scholarly journals Epitope Recognition of a Monoclonal Antibody Raised against a Synthetic Glycerol Phosphate Based Teichoic Acid

2021 ◽  
Author(s):  
Francesca Berni ◽  
Ermioni Kalfopoulou ◽  
Ana M. Gimeno Cardells ◽  
Filippo Carboni ◽  
Daan van der Es ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Teichoic Acid ◽  
Glycerol Phosphate ◽  
Epitope Recognition

scholarly journals The wall content and composition of Bacillus subtilis var. niger grown in a chemostat

1970 ◽  
Vol 118 (3) ◽  
pp. 367-373 ◽  
Author(s):  
D. C. Ellwood
Keyword(s):  
Bacillus Subtilis ◽  
Phosphorus Content ◽  
Teichoic Acid ◽  
Dry Weight ◽  
Growth Conditions ◽  
Glycerol Phosphate ◽  
Limited Growth ◽  
Anionic Polymer ◽  
Rate Analysis ◽  
Teichuronic Acid

Bacillus subtilis var. niger was grown in a chemostat with various growth limitations and at various growth rates. The wall content and composition of the organism grown under these conditions were determined. The wall content, expressed as a percentage of the dry weight of organisms, varied with the growth rate. Analysis of wall samples showed that their composition also varied, particularly with respect to the phosphorus content. Wall samples extracted with trichloroacetic acid under carefully controlled conditions were found to contain various amounts of phosphorus, this being present as a glycerol phosphate polymer containing hexose (glucose and in some cases galactose), i.e. a teichoic aid. Teichoic acids were present in the walls of organisms grown under all conditions except when phosphorus limited growth. Then a different anionic polymer, composed of glucuronic acid and N-acetylgalactosamine (a teichuronic acid), was present. Under the specific growth conditions at pH7.0 and 35°C in a chemostat, teichoic acid and teichuronic acid appeared to be mutually exclusive.

scholarly journals Studies on the linkage between teichoic acid and peptidoglycan in a bacteriophage-resistant mutant of Staphylococcus aureus H

1975 ◽  
Vol 149 (3) ◽  
pp. 637-647 ◽  
Author(s):  
J E Heckels ◽  
A R Archibald ◽  
J Baddiley
Keyword(s):  
Staphylococcus Aureus ◽  
Teichoic Acid ◽  
Resistant Mutant ◽  
Muramic Acid ◽  
Glycerol Phosphate ◽  
Wall Teichoic Acid ◽  
Phosphodiester Linkage ◽  
A Chain

1. In addition to poly(ribitol phosphate) the walls of a bacteriophage-resistant mutant of Staphylococcus aureus H contain glycerol phosphate residues that are not removed on digestion with trypsin or extraction with phenol. 2. The glycerol phosphate is present in a chain, containing three or four glycerol phosphate residues, which is covalently attached to the peptidoglycan through a phosphodiester linkage to muramic acid; this linkage is readily hydrolysed by dilute alkali. 3. The degradative studies described suggest that the poly(ribitol phosphate) chains of the wall teichoic acid may be attached to the wall by linkage to this glycerol phosphate oligomer.

scholarly journals The biosynthesis of the wall teichoic acid in Staphylococcus lactis I3

1968 ◽  
Vol 110 (3) ◽  
pp. 565-571 ◽  
Author(s):  
J Baddiley ◽  
N. L. Blumsom ◽  
L. Julia Douglas
Keyword(s):  
Enzyme System ◽  
Teichoic Acid ◽  
Glycerol Phosphate ◽  
Wall Teichoic Acid ◽  
Enzyme Preparations

1. The biosynthesis of the wall teichoic acid in Staphylococcus lactis I3 was studied. Cell-free particulate enzyme preparations, probably representing fragmented membrane, were isolated and used for the synthesis of polymer. 2. By using appropriately labelled CDP-glycerol and UDP-N-acetylglucosamine it was shown that the former contributes a glycerol phosphate residue and the latter contributes an N-acetylglucosamine 1-phosphate residue to the repeating unit. 3. No polymer was synthesized unless both nucleotides were present, and no other substrates were required. 4. The properties of the enzyme system were studied. 5. Although attempts to fractionate the system failed, the biosynthesis is believed to be complex and its mechanism is considered.

scholarly journals Biosynthesis of the wall teichoic acid in Bacillus licheniformis

1972 ◽  
Vol 127 (1) ◽  
pp. 27-37 ◽  
Author(s):  
I. C. Hancock ◽  
J. Baddiley
Keyword(s):  
Bacillus Licheniformis ◽  
Teichoic Acid ◽  
Chain Extension ◽  
Pulse Labelling ◽  
Glycerol Phosphate ◽  
Wall Teichoic Acid ◽  
Anomeric Configuration ◽  
Glucose Phosphate ◽  
Chemical Studies ◽  
Two Stages

1. The biosynthesis of the wall teichoic acid, poly(glycerol phosphate glucose), has been studied with a particulate membrane preparation from Bacillus licheniformis A.T.C.C. 9945. The precursor CDP-glycerol supplies glycerol phosphate residues, whereas UDP-glucose supplies only glucose to the repeating structure of the polymer. 2. Synthesis proceeds through polyprenol phosphate derivatives, and chemical studies and pulse-labelling techniques show that the first intermediate is the phosphodiester, glucose polyprenol monophosphate. CDP-glycerol donates a glycerol phosphate residue to this to give a second intermediate, (glycerol phosphate glucose phosphate) polyprenol. 3. The glucose residue in the lipid intermediates has the β configuration, and chain extension in the synthesis of polymer occurs by transglycosylation with inversion of anomeric configuration at two stages.

scholarly journals Influence of growth condition on the concentration of potassium in Bacillus subtilis var. niger and its possible relationship to cellular ribonucleic acid, teichoic acid and teichuronic acid

1968 ◽  
Vol 106 (1) ◽  
pp. 237-243 ◽  
Author(s):  
D. W. Tempest ◽  
J. W. Dicks ◽  
D C Ellwood
Keyword(s):  
Bacillus Subtilis ◽  
Dilution Rate ◽  
Cell Walls ◽  
Teichoic Acid ◽  
High Concentration ◽  
Glycerol Phosphate ◽  
Anionic Polymer ◽  
Type Compound ◽  
Acid Type ◽  
Teichuronic Acid

1. Mg2+-limited Bacillus subtilis var. niger, growing in a chemostat in a simple salts medium, contained considerably more potassium and phosphorus than Mg2+-limited Aerobacter aerogenes growing in a similar medium at corresponding dilution rates. 2. Growth of the bacillus in a K+-limited environment did not lower the cellular potassium and phosphorus contents, the molar proportions of cell-bound magnesium, potassium, RNA (as nucleotide) and phosphorus being approximately constant at 1:13:5:13 (compared with 1:4:5:8 in Mg2+-limited or K+-limited A. aerogenes). 3. Growth of B. subtilis in a phosphate-limited environment caused the cellular phosphorus content to be lowered to a value similar to that of Mg2+-limited A. aerogenes, but the potassium content was not correspondingly lowered; the molar potassium:magnesium ratio varied from 14 to 17 with changes in dilution rate from 0·4 to 0·1hr.−1. 4. Whereas over 70% of the cell-bound phosphorus of Mg2+-limited or K+-limited A. aerogenes was contained in the nucleic acids, these polymers accounted for less than 50% of the phosphorus present in similarly limited B. subtilis; much phosphorus was present in the walls of the bacilli, bound in a teichoic acid-type compound composed of glycerol phosphate and glucose (but no alanine). 5. Phosphate-limited B. subtilis cell walls (from organisms grown at a dilution rate of 0·2hr.−1) contained little phosphorus and no detectable amounts of teichoic acid, but 40% of the cell-wall dry weight could be accounted for by a teichuronic acid-type compound; this contained a glucuronic acid and galactosamine, neither of which could be detected in the walls of Mg2+-limited B. subtilis grown at a corresponding rate. 6. It is suggested that the high concentration of potassium in growing B. subtilis (compared with A. aerogenes) results from the presence of large amounts of anionic polymer (teichoic acid or teichuronic acid) in the bacillus cell walls.

scholarly journals The glycerol teichoic acid of walls of Staphylococcus lactis I3

1968 ◽  
Vol 110 (3) ◽  
pp. 543-557 ◽  
Author(s):  
A. R. Archibald ◽  
J Baddiley ◽  
D. Button
Keyword(s):  
Trichloroacetic Acid ◽  
Teichoic Acid ◽  
Glycerol Phosphate ◽  
Polymeric Structure ◽  
Phosphodiester Group ◽  
Mild Acid

1. The teichoic acid from walls of Staphylococcus lactis I3 was isolated by extraction with trichloroacetic acid and shown to contain glycerol, N-acetylglucosamine, phosphate and d-alanine in the molecular proportions 1:1:2:1. The alanine is attached to the polymer through ester linkages. 2. Hydrolysis with acid gave alanine, glucosamine and glycerol diphosphates. Under mild acid conditions a repeating unit was produced; this consists of glycerol diphosphate joined through a phosphodiester group to N-acetylglucosamine. 3. Hydrolysis with alkali gave glycerol diphosphates, saccharinic acid and two phosphodiesters containing glucosamine whose structures were elucidated; these both contain glucosamine 1-phosphate, and N-acetylglucosamine 1-phosphate was isolated by a degradative procedure. 4. The unusual properties of the teichoic acid are explained by a polymeric structure in which N-acetylglucosamine 1-phosphate is attached through its phosphate to glycerol phosphate. 5. The biosynthetic implications of this structure are discussed.

scholarly journals Teichoic acid synthesis in Bacillus stearothermophilus

1974 ◽  
Vol 138 (3) ◽  
pp. 525-535 ◽  
Author(s):  
L. D. Kennedy
Keyword(s):  
Alkaline Hydrolysis ◽  
Bacillus Stearothermophilus ◽  
De Novo ◽  
Enzyme System ◽  
Teichoic Acid ◽  
Periodate Oxidation ◽  
Acid Synthesis ◽  
Pulse Labelling ◽  
Glycerol Phosphate ◽  
Enzyme Preparations

1. Particulate enzyme preparations obtained from Bacillus stearothermophilus B65 by digestion with lysozyme were shown to catalyse teichoic acid synthesis. With CDP-glycerol as sole substrate the preparations synthesized 1,3-poly(glycerol phosphate). It was characterized by alkaline hydrolysis, by glucosylation to the alkali-stable 2-glucosyl-1,3-poly(glycerol phosphate) with excess of UDP-glucose and a Bacillus subtilis Marburg enzyme system, by degradation of this latter product with 60%HF and periodate oxidation of the resulting glucosylglycerol. The specificity of the B. subtilis system previously reported (Glaser & Burger, 1964), was confirmed in the present work. 2. Pulse-labelling experiments, followed by periodate oxidation of the product and isolation of formaldehyde from the glycerol terminus of the polymer, showed that the B. stearothermophilus enzyme system transferred glycerol phosphate units to the glycerol end of the chain. The transfer reaction was irreversible. It was not determined if these poly(glycerol phosphate) chains were synthesized de novo, but it was shown that the newly synthesized oligomers were bound to much larger molecules. 3. When the B. stearothermophilus enzyme system was supplied with both CDP-glycerol and UDP-glucose, 1-glucosyl-2,3-poly(glycerol phosphate) was synthesized in addition to the 1,3-isomer. The former polymer was characterized by acid and alkaline hydrolysis, degradation with HF and periodate oxidation of the resulting glucosylglycerol, and periodate oxidation of the intact polymer followed by mild acid hydrolysis. This latter procedure removed the glucose substituents without disrupting the poly(glycerol phosphate) chain. 4. The poly(glycerol phosphate) isomers were distinguished by glucosylation with the B. subtilis enzymes and alkaline hydrolysis, the 2,3-isomer remaining alkali-labile. The proportion of 2,3-poly(glycerol phosphate) in the product increased with increasing amounts of UDP-glucose in the incubation mixture, but the total glycerol phosphate incorporated into products remained constant. It is suggested that the synthetic pathways of the two poly(glycerol phosphate) species may share a rate-limiting step.

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