Fluorescence and Atomic Force Microscopy Imaging of Wall Teichoic Acids in <i>Lactobacillus plantarum</i>

André, Guillaume; Deghorain, Marie; Bron, Peter A.; Swam, Iris I. van; Kleerebezem, Michiel; Hols, Pascal; Dufrêne, Yves F.

doi:10.1021/cb1003509

Cited by 74 publications

(81 citation statements)

References 45 publications

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“…Construction of mutants lacking WTAs has been successfully demonstrated in B. subtilis, S. aureus and Lb. plantarum (D'Elia et al, 2006a, b;Andre et al, 2011), showing that WTA is not indispensable to the growth of bacterial cells. Hence, the variety and uniformity of WTA glucosylation would have an impact on survival and adaptation to different environments.…”

Section: Discussionmentioning

confidence: 99%

Determination of strain-specific wall teichoic acid structures in Lactobacillus plantarum reveals diverse α-d-glucosyl substitutions and high structural uniformity of the repeating units

et al. 2012

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The structural diversity of wall teichoic acid (WTA) was investigated using biochemical and NMR analyses among 19 strains of Lactobacillus plantarum, of which seven were previously established to contain a glycerol-type backbone, whereas the remaining 12 strains possess ribitol-containing WTA. Despite the fact that the WTAs consisted of identical components, namely phosphoric acid, alditol (glycerol or ribitol) and glucose, comparative analysis of the 1 H and 13 C NMR spectra indicated the presence of six different structures, based on the observed differences in the anomeric signals of glucose residues. To determine the six WTA structures, their repeating units were prepared by alkaline hydrolysis, followed by fractionation on HPLC, and analysis by NMR spectroscopy using synthetic molecules as a reference. The structures of the six isolates were established as 1-a-D-glucosyl-sn-glycerol 3-phosphate, 1-a-D-kojibiosyl-sn-glycerol 3-phosphate, 1-a-D-nigerosyl-sn-glycerol 3-phosphate, 4-a-D-kojibiosylribitol 1-phosphate and 1,5-linked di-(2,4-di-a-D-glucosylribitol) phosphate. The backbone structures appeared to be 3,69-linked poly(1-a-D-glucosyl-sn-glycerol phosphate) for the glycerol-type WTA and 1,5-linked poly(ribitol phosphate) for the ribitol-containing WTA. Moreover, in the analysis of the alkaline hydrolysates on HPLC, only single structures of repeating units were released from each WTA, indicating the high structural uniformity of the WTA in each strain. Notably, analyses of lipoteichoic acid isolated from representative strains harbouring the six different WTAs revealed the universal presence of a 1,3-linked poly(glycerol phosphate) chain, substituted at C-2 of the glycerol residues with glucose residues. These findings provide fundamental information on WTA structural variability in Lb. plantarum, which seems likely to play a pivotal role in the physiology of this bacterial species.

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Section: Discussionmentioning

confidence: 99%

Determination of strain-specific wall teichoic acid structures in Lactobacillus plantarum reveals diverse α-d-glucosyl substitutions and high structural uniformity of the repeating units

et al. 2012

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“…Microscopy analyses were performed using an Axio observer Z1 inverted microscope (Carl Zeiss). FM4-64 (Molecular Probes, Leiden, The Netherlands) and DAPI (4Ј,6-diamidino-2-phenylindole) (Sigma, Bornem, Belgium) staining was performed as previously reported (1). Analyses of micrographies were performed using the AxioVision 4.8. software (Carl Zeiss).…”

Section: Methodsmentioning

confidence: 99%

Identification of the Amidotransferase AsnB1 as Being Responsible for meso -Diaminopimelic Acid Amidation in Lactobacillus plantarum Peptidoglycan

et al. 2011

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The peptidoglycan (PG) of Lactobacillus plantarum contains amidated meso-diaminopimelic acid (mDAP). The functional role of this PG modification has never been characterized in any bacterial species, except for its impact on PG recognition by receptors of the innate immune system. In silico analysis of loci carrying PG biosynthesis genes in the L. plantarum genome revealed the colocalization of the murE gene, which encodes the ligase catalyzing the addition of mDAP to UDP-N-muramoyl-D-glutamate PG precursors, with asnB1, which encodes a putative asparagine synthase with an N-terminal amidotransferase domain. By gene disruption and complementation experiments, we showed that asnB1 is the amidotransferase involved in mDAP amidation. PG structural analysis revealed that mDAP amidation plays a key role in the control of the L,D-carboxypeptidase DacB activity. In addition, a mutant strain with a defect in mDAP amidation is strongly affected in growth and cell morphology, with filamentation and cell chaining, while a DacB-negative strain displays a phenotype very similar to that of a wild-type strain. These results suggest that mDAP amidation may play a critical role in the control of the septation process.Peptidoglycan (PG) is a heteropolymer of glycan strands cross-linked by peptidic stems most often between the fourth and the third amino acids of the donor and the acceptor stem, respectively. The composition of this peptidic stem can vary from one bacterial species to another and, in Lactobacillus plantarum, is composed of L-Ala, D-Glu, meso-diaminopimelic acid (mDAP), and D-Ala. D-Lactic acid is found as the last moiety of the peptidic stem in PG precursors (5).Different amino acids found in PG have been reported as amidated in various species: D-Glu into D-iso-Gln (5, 9, 14, 21), mDAP into amidated mDAP (3,5,21), and D-Asp into D-Asn in species possessing a D-Asp as a cross bridge. Among these PG modifications, only the D-Asp amidation was characterized in Lactococcus lactis (23). D-Asp amidation is catalyzed by an asparagine synthase, AsnH, which is involved in autolysis control and resistance to cationic peptides (23).We have recently shown that both D-Glu and mDAP are highly amidated (100% and 94%, respectively) in L. plantarum (5). The functional role of these amidations remains poorly understood, except for their impact on PG recognition by the mammalian host innate immune system (2, 11, 14). As a typical prokaryotic structure, PG is sensed by pattern recognition receptors involved in bacterial detection (e.g., Nod1, Nod2, and Toll-like receptor 2[TLR2]), and PG modifications were shown to affect this process (2, 6, 11, 12). Unamidated mDAP is essential for Nod1 detection (2), whereas amidated mDAP was reported to modulate the recognition by TLR2 (11). Despite these important immunomodulatory properties, the genetic determinants of mDAP amidation have never been described, and the functional role of mDAP amidation in bacterial physiology remains completely unexplored.In this study, we identify the first mDAP...

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“…In recent years, atomic-force microscopy (AFM) has provided valuable information on the structural, adhesive, and mechanical properties of the cell wall in numerous biological samples (17)(18)(19)(20)(21)(22)(23)(24). AFM can be used not only to obtain topological information on the cell surface and the organization of the peptidoglycan (20,22,(24)(25)(26), but it is also a powerful tool for quantitative studies of physical properties of the surface, such as the elasticity of the bacterial cell wall (27)(28)(29)(30)(31)(32)(33)(34).…”

Section: Introductionmentioning

confidence: 99%

Reduction of the Peptidoglycan Crosslinking Causes a Decrease in Stiffness of the Staphylococcus aureus Cell Envelope

Loskill

Pereira

Jung

et al. 2014

Biophysical Journal

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We have used atomic-force microscopy (AFM) to probe the effect of peptidoglycan crosslinking reduction on the elasticity of the Staphylococcus aureus cell wall, which is of particular interest as a target for antimicrobial chemotherapy. Penicillin-binding protein 4 (PBP4) is a nonessential transpeptidase, required for the high levels of peptidoglycan crosslinking characteristic of S. aureus. Importantly, this protein is essential for β-lactam resistance in community-acquired, methicillin-resistant S. aureus (MRSA) strains but not in hospital-acquired MRSA strains. Using AFM in a new mode for recording force/distance curves, we observed that the absence of PBP4, and the concomitant reduction of the peptidoglycan crosslinking, resulted in a reduction in stiffness of the S. aureus cell wall. Importantly, the reduction in cell wall stiffness in the absence of PBP4 was observed both in community-acquired and hospital-acquired MRSA strains, indicating that high levels of peptidoglycan crosslinking modulate the overall structure and mechanical properties of the S. aureus cell envelope in both types of clinically relevant strains. Additionally, we were able to show that the applied method enables the separation of cell wall properties and turgor pressure.

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Fluorescence and Atomic Force Microscopy Imaging of Wall Teichoic Acids in Lactobacillus plantarum

Cited by 74 publications

References 45 publications

Determination of strain-specific wall teichoic acid structures in Lactobacillus plantarum reveals diverse α-d-glucosyl substitutions and high structural uniformity of the repeating units

Determination of strain-specific wall teichoic acid structures in Lactobacillus plantarum reveals diverse α-d-glucosyl substitutions and high structural uniformity of the repeating units

Identification of the Amidotransferase AsnB1 as Being Responsible for meso -Diaminopimelic Acid Amidation in Lactobacillus plantarum Peptidoglycan

Reduction of the Peptidoglycan Crosslinking Causes a Decrease in Stiffness of the Staphylococcus aureus Cell Envelope

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