scholarly journals The first biantennary bacterial secondary cell wall polymer and its influence on S-layer glycoprotein assembly

2002 ◽  
Vol 368 (2) ◽  
pp. 483-494 ◽  
Author(s):  
Christian STEINDL ◽  
Christina SCHÄFFER ◽  
Thomas WUGEDITSCH ◽  
Michael GRANINGER ◽  
Irena MATECKO ◽  
...  
Keyword(s):  
Cell Wall ◽  
Self Assembly ◽  
Secondary Cell Wall ◽  
Water Soluble ◽  
Bacterial Cells ◽  
Cell Wall Polymer ◽  
Thaw Cycle ◽  
Secondary Cell Wall Polymer

The cell surface of Aneurinibacillus thermoaerophilus DSM 10155 is covered with a square surface (S)-layer glycoprotein lattice. This S-layer glycoprotein, which was extracted with aqueous buffers after a freeze—thaw cycle of the bacterial cells, is the only completely water-soluble S-layer glycoprotein to be reported to date. The purified S-layer glycoprotein preparation had an overall carbohydrate content of 19%. Detailed chemical investigations indicated that the S-layer O-glycans of previously established structure accounted for 13% of total glycosylation. The remainder could be attributed to a peptidoglycan-associated secondary cell wall polymer. Structure analysis was performed using purified secondary cell wall polymer—peptidoglycan complexes. NMR spectroscopy revealed the first biantennary secondary cell wall polymer from the domain Bacteria, with the structure α-d-GlcpNAc-(1→3)-β-d-ManpNAc-(1→4)-β-d-GalpNAc-(1→3)-α-d-GlcpNAc-(1→3)-β-d-ManpNAc-(1→4)-β-d-GalpNAc-(1→3)-α-d-GlcpNAc-(1→4)-[α-d-GlcpNAc-(1→3)-β-d-ManpNAc-(1→4)-β-d-GalpNAc-(1→3)-α-d-GlcpNAc-(1→3)-β-d-ManpNAc-(1→4)-β-d-GalpNAc-(1→3)-α-d-GlcpNAc-(1→3)]-β-d-ManpNAc-(1→3)-α-d-GlcpNAc-(1→3)-β-d-ManpNAc-(1→3)-α-d-GlcpNAc-(1→3)-α-d-GlcpNAc-(1→O)-PO2--O-PO2--(O→6)-MurNAc- (where MurNAc is N-acetylmuramic acid). The neutral polysaccharide is linked via a pyrophosphate bond to the C-6 atom of every fourth N-acetylmuramic acid residue, in average, of the A1γ-type peptidoglycan. In vivo, the biantennary polymer anchored the S-layer glycoprotein very effectively to the cell wall, probably due to the doubling of motifs for a proposed lectin-like binding between the polymer and the N-terminus of the S-layer protein. When the cellular support was removed during S-layer glycoprotein isolation, the co-purified polymer mediated the solubility of the S-layer glycoprotein in vitro. Initial crystallization experiments performed with the soluble S-layer glycoprotein revealed that the assembly property could be restored upon dissociation of the polymer by the addition of poly(ethylene glycols). The formed two-dimensional crystalline S-layer self-assembly products exhibited the same lattice symmetry as observed on intact bacterial cells.

scholarly journals Influence of the Secondary Cell Wall Polymer on the Reassembly, Recrystallization, and Stability Properties of the S-Layer Protein from Bacillus stearothermophilus PV72/p2

1998 ◽  
Vol 180 (16) ◽  
pp. 4146-4153 ◽  
Author(s):  
Margit Sára ◽  
Christine Dekitsch ◽  
Harald F. Mayer ◽  
Eva M. Egelseer ◽  
Uwe B. Sleytr
Keyword(s):  
Cell Wall ◽  
Self Assembly ◽  
Secondary Cell Wall ◽  
Bacillus Stearothermophilus ◽  
Guanidine Hydrochloride ◽  
Terminal Region ◽  
Water Soluble ◽  
Outer Face ◽  
Cell Wall Polymer ◽  
Secondary Cell Wall Polymer

ABSTRACT The high-molecular-weight secondary cell wall polymer (SCWP) fromBacillus stearothermophilus PV72/p2 is mainly composed ofN-acetylglucosamine (GlcNAc) andN-acetylmannosamine (ManNAc) and is involved in anchoring the S-layer protein via its N-terminal region to the rigid cell wall layer. In addition to this binding function, the SCWP was found to inhibit the formation of self-assembly products during dialysis of the guanidine hydrochloride (GHCl)-extracted S-layer protein. The degree of assembly (DA; percent assembled from total S-layer protein) that could be achieved strongly depended on the amount of SCWP added to the GHCl-extracted S-layer protein and decreased from 90 to 10% when the concentration of the SCWP was increased from 10 to 120 μg/mg of S-layer protein. The SCWP kept the S-layer protein in the water-soluble state and favored its recrystallization on solid supports such as poly-l-lysine-coated electron microscopy grids. Derived from the orientation of the base vectors of the oblique S-layer lattice, the subunits had bound with their charge-neutral outer face, leaving the N-terminal region with the polymer binding domain exposed to the ambient environment. From cell wall fragments about half of the S-layer protein could be extracted with 1 M GlcNAc, indicating that the linkage type between the S-layer protein and the SCWP could be related to that of the lectin-polysaccharide type. Interestingly, GlcNAc had an effect on the in vitro self-assembly and recrystallization properties of the S-layer protein that was similar to that of the isolated SCWP. The SCWP generally enhanced the stability of the S-layer protein against endoproteinase Glu-C attack and specifically protected a potential cleavage site in position 138 of the mature S-layer protein.

In vitro and in vivo polysaccharides assembly: ultrastructural and cytochemical study of growing plant cell wall components.

1978 ◽  
Vol 36 (2) ◽  
pp. 434-435
Author(s):  
D. Reis ◽  
B. Vian ◽  
J. C. Roland
Keyword(s):  
Cell Wall ◽  
Self Assembly ◽  
Plant Cell Wall ◽  
Higher Plants ◽  
Three Dimensional ◽  
Cytochemical Study ◽  
Wall Components

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.

scholarly journals Non-Lytic Antibacterial Peptides That Translocate Through Bacterial Membranes to Act on Intracellular Targets

2019 ◽  
Vol 20 (19) ◽  
pp. 4877 ◽  
Author(s):  
Marlon H. Cardoso ◽  
Beatriz T. Meneguetti ◽  
Bruna O. Costa ◽  
Danieli F. Buccini ◽  
Karen G. N. Oshiro ◽  
...  
Keyword(s):  
Cell Wall ◽  
Protein Synthesis ◽  
Pathogenic Bacteria ◽  
Biological Properties ◽  
Antibacterial Peptides ◽  
Bacterial Cells ◽  
Bacterial Membranes ◽  
Intracellular Targets

The advent of multidrug resistance among pathogenic bacteria has attracted great attention worldwide. As a response to this growing challenge, diverse studies have focused on the development of novel anti-infective therapies, including antimicrobial peptides (AMPs). The biological properties of this class of antimicrobials have been thoroughly investigated, and membranolytic activities are the most reported mechanisms by which AMPs kill bacteria. Nevertheless, an increasing number of works have pointed to a different direction, in which AMPs are seen to be capable of displaying non-lytic modes of action by internalizing bacterial cells. In this context, this review focused on the description of the in vitro and in vivo antibacterial and antibiofilm activities of non-lytic AMPs, including indolicidin, buforin II PR-39, bactenecins, apidaecin, and drosocin, also shedding light on how AMPs interact with and further translocate through bacterial membranes to act on intracellular targets, including DNA, RNA, cell wall and protein synthesis.

scholarly journals High-Affinity Interaction between the S-Layer Protein SbsC and the Secondary Cell Wall Polymer of Geobacillus stearothermophilus ATCC 12980 Determined by Surface Plasmon Resonance Technology

2007 ◽  
Vol 189 (19) ◽  
pp. 7154-7158 ◽  
Author(s):  
Judith Ferner-Ortner ◽  
Christoph Mader ◽  
Nicola Ilk ◽  
Uwe B. Sleytr ◽  
Eva M. Egelseer
Keyword(s):  
Amino Acids ◽  
Cell Wall ◽  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Secondary Cell Wall ◽  
Geobacillus Stearothermophilus ◽  
Binding Behavior ◽  
Cell Wall Polymer ◽  
Secondary Cell Wall Polymer

ABSTRACT Surface plasmon resonance studies using C-terminal truncation forms of the S-layer protein SbsC (recombinant SbsC consisting of amino acids 31 to 270 [rSbsC31-270] and rSbsC31-443) and the secondary cell wall polymer (SCWP) isolated from Geobacillus stearothermophilus ATCC 12980 confirmed the exclusive responsibility of the N-terminal region comprising amino acids 31 to 270 for SCWP binding. Quantitative analyses indicated binding behavior demonstrating low, medium, and high affinities.

scholarly journals Structural and Functional Analyses of the Secondary Cell Wall Polymer of Bacillus sphaericus CCM 2177 That Serves as an S-Layer-Specific Anchor

1999 ◽  
Vol 181 (24) ◽  
pp. 7643-7646 ◽  
Author(s):  
Nicola Ilk ◽  
Paul Kosma ◽  
Michael Puchberger ◽  
Eva M. Egelseer ◽  
Harald F. Mayer ◽  
...  
Keyword(s):  
Cell Wall ◽  
Hydrofluoric Acid ◽  
Secondary Cell Wall ◽  
Bacillus Sphaericus ◽  
Nuclear Magnetic Resonance Analysis ◽  
Terminal Part ◽  
Resonance Analysis ◽  
Cell Wall Polymer ◽  
Functional Analyses ◽  
Secondary Cell Wall Polymer

ABSTRACT Sacculi of Bacillus sphaericus CCM 2177 contain a secondary cell wall polymer which was completely extracted with 48% hydrofluoric acid. Nuclear magnetic resonance analysis showed that the polymer is composed of repeating units, as follows: →3)-[4,6-O-(1-carboxyethylidene)]∼0.5-β-d-ManpNAc-(1→4)-β-d-GlcpNAc-(1→. The N-terminal part of the S-layer protein carrying S-layer homologous motifs recognizes this polymer as a binding site.

scholarly journals Structural basis of peptidoglycan endopeptidase regulation

2020 ◽  
Vol 117 (21) ◽  
pp. 11692-11702 ◽  
Author(s):  
Jung-Ho Shin ◽  
Alan G. Sulpizio ◽  
Aaron Kelley ◽  
Laura Alvarez ◽  
Shannon G. Murphy ◽  
...  
Keyword(s):  
Cell Wall ◽  
Structural Basis ◽  
Bacterial Cells ◽  
Cell Wall Degradation ◽  
Open Conformation ◽  
Poor Understanding ◽  
A Cell ◽  
Inhibitory Domain

Most bacteria surround themselves with a cell wall, a strong meshwork consisting primarily of the polymerized aminosugar peptidoglycan (PG). PG is essential for structural maintenance of bacterial cells, and thus for viability. PG is also constantly synthesized and turned over; the latter process is mediated by PG cleavage enzymes, for example, the endopeptidases (EPs). EPs themselves are essential for growth but also promote lethal cell wall degradation after exposure to antibiotics that inhibit PG synthases (e.g., β-lactams). Thus, EPs are attractive targets for novel antibiotics and their adjuvants. However, we have a poor understanding of how these enzymes are regulated in vivo, depriving us of novel pathways for the development of such antibiotics. Here, we have solved crystal structures of the LysM/M23 family peptidase ShyA, the primary EP of the cholera pathogenVibrio cholerae. Our data suggest that ShyA assumes two drastically different conformations: a more open form that allows for substrate binding and a closed form, which we predicted to be catalytically inactive. Mutations expected to promote the open conformation caused enhanced activity in vitro and in vivo, and these results were recapitulated in EPs from the divergent pathogensNeisseria gonorrheaeandEscherichia coli. Our results suggest that LysM/M23 EPs are regulated via release of the inhibitory Domain 1 from the M23 active site, likely through conformational rearrangement in vivo.

scholarly journals The S-Layer Proteins of Two Bacillus stearothermophilus Wild-Type Strains Are Bound via Their N-Terminal Region to a Secondary Cell Wall Polymer of Identical Chemical Composition

1998 ◽  
Vol 180 (6) ◽  
pp. 1488-1495 ◽  
Author(s):  
Eva Maria Egelseer ◽  
Karl Leitner ◽  
Marina Jarosch ◽  
Christoph Hotzy ◽  
Sonja Zayni ◽  
...  
Keyword(s):  
Cell Wall ◽  
Secondary Cell Wall ◽  
Bacillus Stearothermophilus ◽  
Proteolytic Cleavage ◽  
Terminal Region ◽  
Wild Type ◽  
Cell Wall Polymer ◽  
Secondary Cell Wall Polymer ◽  
Type Strains ◽  
Identical Chemical Composition

ABSTRACT Two Bacillus stearothermophilus wild-type strains were investigated regarding a common recognition and binding mechanism between the S-layer protein and the underlying cell envelope layer. The S-layer protein from B. stearothermophilusPV72/p6 has a molecular weight of 130,000 and assembles into a hexagonally ordered lattice. The S-layer from B. stearothermophilus ATCC 12980 shows oblique lattice symmetry and is composed of subunits with a molecular weight of 122,000. Immunoblotting, peptide mapping, N-terminal sequencing of the whole S-layer protein from B. stearothermophilus ATCC 12980 and of proteolytic cleavage fragments, and comparison with the S-layer protein from B. stearothermophilus PV72/p6 revealed that the two S-layer proteins have identical N-terminal regions but no other extended structurally homologous domains. In contrast to the heterogeneity observed for the S-layer proteins, the secondary cell wall polymer isolated from peptidoglycan-containing sacculi of the different strains showed identical chemical compositions and comparable molecular weights. The S-layer proteins could bind and recrystallize into the appropriate lattice type on native peptidoglycan-containing sacculi from both organisms but not on those extracted with hydrofluoric acid, leading to peptidoglycan of the A1γ chemotype. Affinity studies showed that only proteolytic cleavage fragments possessing the complete N terminus of the mature S-layer proteins recognized native peptidoglycan-containing sacculi as binding sites or could associate with the isolated secondary cell wall polymer, while proteolytic cleavage fragments missing the N-terminal region remained unbound. From the results obtained in this study, it can be concluded that S-layer proteins from B. stearothermophilus wild-type strains possess an identical N-terminal region which is responsible for anchoring the S-layer subunits to a secondary cell wall polymer of identical chemical composition.

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