Structural characterization of the lipoteichoic acid isolated from Staphylococcus sciuri W620

Duda, Katarzyna; Petersen, Sandra; Holst, Otto

doi:10.1016/j.carres.2016.04.026

Cited by 3 publications

(5 citation statements)

References 23 publications

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“…To define, if LTA serves as a cell wall ligand for the L. buchneri CD034 S-layer, CWGs were extracted with n -butanol from disrupted L. buchneri CD034 cells whose S-layer had been stripped-off before, and further separated by hydrophobic interaction chromatography (HIC) ( Morath et al 2001 ; Duda et al 2016 ), yielding two pools. Fractions eluting from the HIC column isocratically with 15% n -propanol in 0.1 M ammonium acetate (pH 4.7) represented pool I and those eluting in the HIC gradient between 45% and 65% n -propanol in 0.1 M ammonium acetate (pH 4.7) were combined to pool II.…”

Section: Resultsmentioning

confidence: 99%

“…For extraction of CWGs, disrupted L. buchneri cells were stirred with an equal volume of n -butanol (LiChrosolv, Merck, Darmstadt, Germany) at 22°C for 1 h ( Morath et al 2001 ). After centrifugation (7500 × g , 30 min), the aqueous phase containing CWGs was collected and dialyzed for 4 d against distilled water and lyophilized ( Duda et al 2016 ). Cell wall glycopolymers were purified from that phase by hydrophobic interaction chromatography (HIC) on a HiPrep column (16 mm × 100 mm, bed volume 20 mL) of octyl-Sepharose (GE Healthcare, Little Chalfont, UK) to separate LTA from co-extracted CWGs ( Morath et al 2001 ).…”

Section: Methodsmentioning

confidence: 99%

“…The components of L. buchneri CWGs were determined by methanolysis with 0.5 M HCl/methanol (MeOH) at 85°C for 45 min, followed by acetylation using acetic anhydride and pyridine (1:1 [v/v]) at 85°C for 10 min and detection by GLC and GLC-mass spectrometry (Agilent Technologies 7890 A gas chromatograph [Agilent Technologies, Wilmington, DE] equipped with a dimethylpolysiloxane column [Agilent, HP Ultra 1, 12 m × 0.2 mm × 0.33 μm film thickness] and a 5975 C series MSD detector with electron impact ionization [EI] mode) under autotune condition at 70 eV. The temperature program was 70°C for 1.5 min, then 60°C/min to 110°C and 5°C/min to 320°C ( Duda et al 2016 ).…”

Section: Methodsmentioning

confidence: 99%

“…The temperature in GLC was 150°C for 3 min, and then increased with 3°C/min to 320°C. For quantification, xylose was used as internal standard and the sugars were identified by use of authentic standards ( Duda et al 2016 ). The absolute configuration of Glc and Gal was determined by GLC of the acetylated O -[( S )-2-butyl] glycoside after methanolysis (2 M HCl/MeOH, 85°C, 45 min), butanolysis (2 M HCl/( S )-2-BuOH, 65°C, 4 h) and peracetylation (85°C, 10 min), by comparison with authentic standards ( Gerwig et al 1979 ).…”

Section: Methodsmentioning

confidence: 99%

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Lipoteichoic acid mediates binding of a Lactobacillus S-layer protein

Bönisch

Anzengruber

et al. 2018

Glycobiology

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The Gram-positive lactic acid bacterium Lactobacillus buchneri CD034 is covered by a two-dimensional crystalline, glycoproteinaceous cell surface (S-) layer lattice. While lactobacilli are extensively exploited as cell surface display systems for applied purposes, questions about how they stick their cell wall together are remaining open. This also includes the identification of the S-layer cell wall ligand. In this study, lipoteichoic acid was isolated from the L. buchneri CD034 cell wall as a significant fraction of the bacterium’s cell wall glycopolymers, structurally characterized and analyzed for its potential to mediate binding of the S-layer to the cell wall. Combined component analyses and 1D- and 2D-nuclear magnetic resonance spectroscopy (NMR) revealed the lipoteichoic acid to be composed of on average 31 glycerol-phosphate repeating units partially substituted with α-d-glucose, and with an α-d-Galp(1→2)-α-d-Glcp(1→3)−1,2-diacyl-sn-Gro glycolipid anchor. The specificity of binding between the L. buchneri CD034 S-layer protein and purified lipoteichoic acid as well as their interaction force of about 45 pN were obtained by single-molecule force spectroscopy; this value is in the range of typical ligand–receptor interactions. This study sheds light on a functional implication of Lactobacillus cell wall architecture by showing direct binding between lipoteichoic acid and the S-layer of L. buchneri CD034.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Lipoteichoic acid mediates binding of a Lactobacillus S-layer protein

Bönisch

Anzengruber

et al. 2018

Glycobiology

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“…Second, although mecA1 is known to encode a protein with a molecular weight of 75 kDa (Genbank no. Y13052) ( Duda et al, 2016 ), the expression product of mecA1 has been detected with a higher molecular weight of 84 kDa by using the antibody of PBP2a (encoded by mecA ) ( Wu et al, 2001 ; Rolo et al, 2017 ; Alijani et al, 2019 ). This suggests that MecA (PBP2a) antibody may not be effective in detecting the expression of mecA1 .…”

Section: Discussionmentioning

confidence: 99%

Inducible Resistance to β-Lactams in Oxacillin-Susceptible mecA1-Positive Staphylococcus sciuri Isolated From Retail Pork

Cai

et al. 2021

Front. Microbiol.

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Most isolated strains of Staphylococcus sciuri contain mecA1, the evolutionary origin of mecA, but are sensitive to β-lactams (OS-MRSS, oxacillin-susceptible mecA1-positive S. sciuri). In order to improve the efficacy of antibiotic treatment, it is important to clarify whether the resistance of OS-MRSS to β-lactams is an inducible phenotype. In this study, three OS-MRSS strains with oxacillin MIC = 1 μg/ml were isolated from 29 retail pork samples. The resistance of OS-MRSS to β-lactams (MIC > 256 μg/ml) was found to be induced by oxacillin, and the induced resistance was observed to remain stable within a certain period of time. Interestingly, the induced β-lactam resistance was not caused by mecA1, heterogeneous resistance, or any genetic mutation, but mainly due to increased wall teichoic acid (WTA) synthesis that thickened the cell wall. The induced strains also showed slower growth rate, as well as decreased adhesion ability and biofilm thickness. These phenotypes were found to be achieved through altered gene expression in associated pathways, such as the citrate cycle and pentose phosphate pathway. The results challenge the traditional antibiotic sensitivity test. In the presence of β-lactam antibiotics, OS-MRSS that was initially sensitive to β-lactams was observed to gradually develop β-lactam resistance in several days. This often-neglected phenomenon in antibiotic sensitivity tests requires further research attention.

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Structural, Biosynthetic, and Serological Cross-Reactive Elucidation of Capsular Polysaccharides from Streptococcus pneumoniae Serogroup 16

Duda

Elverdal³

et al. 2019

J Bacteriol

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Capsular polysaccharides (CPS) are crucial virulence factors of Streptococcus pneumoniae. The previously unknown CPS structures of the pneumococcal serogroup 16 (serotypes 16F and 16A) were thoroughly elucidated by nuclear magnetic resonance (NMR) spectroscopy and verified by chemical analysis. The following repeat unit structures were determined: 16F, -3)-α-l-Rhap-[4-P-1-Gro]-(1-3)-α-d-Glcp-[(6-P-1)-Gro]-(1-3)-β-l-Rhap-[2-OAc]-(1-4)-β-d-Glcp-(1-; 16A, -3)-β-d-Galf-[2-OAc (70%)]-(1-3)-α-l-Rhap-(1-2)-α-l-Rhap-(1-3)-α-d-Galp-[(6-P-1)-Gro]-(1-3)-β-d-Galp-(1-4)-β-d-Glcp-(1- (OAc, O-acetyl substitution; P-1-Gro, glycerol-1-phosphate substitution). A further analysis of CPS biosynthesis of serotypes 16F and 16A, in conjunction with published cps gene bioinformatics analysis and structures of related serotypes, revealed presumable specific function of glycosyltransferase, acetyltransferase, phosphotransferase, and polymerase. The functions of glycosyltransferases WcxN and WcxT were proposed for the first time, and they were assigned to catalyze linkage of α-l-Rhap-(1-3)-α-d-Glcp and α-l-Rhap-(1-2)-α-l-Rhap, respectively. Furthermore, since serotype 16F was genetically close to serogroup 28, cross-reactions between serogroup 16 and serogroup 28 were studied using diagnostic antisera, which provided further understanding of antigenic properties of CPS and diagnostic antisera. Interestingly, serotype 16F cross-reacted with factor antisera 28b and 11c. Meanwhile, serotype 16A cross-reacted with factor antiserum 11c. IMPORTANCE The vaccine pressure against Streptococcus pneumoniae could result in a change of prevalence in carriage and invasive serotypes. As such, it is necessary to monitor the distribution to achieve successful vaccination of the population, and similarly, it is important to increase the knowledge of even the currently less prevalent serotypes. The CPS are vital for the virulence of the pathogen, and antigenic properties of CPS are based on the structure. Consequently, a better understanding of the structure, biosynthesis, and serology of the capsular polysaccharides can be of great importance toward developing future diagnostic tools and vaccines.

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Structural characterization of the lipoteichoic acid isolated from Staphylococcus sciuri W620

Cited by 3 publications

References 23 publications

Lipoteichoic acid mediates binding of a Lactobacillus S-layer protein

Lipoteichoic acid mediates binding of a Lactobacillus S-layer protein

Inducible Resistance to β-Lactams in Oxacillin-Susceptible mecA1-Positive Staphylococcus sciuri Isolated From Retail Pork

Structural, Biosynthetic, and Serological Cross-Reactive Elucidation of Capsular Polysaccharides from Streptococcus pneumoniae Serogroup 16

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