Interaction of the Staphylococcin-like Peptide Pep 5 with Cell Walls and Isolated Cell Wall Components of Gram-Positive Bacteria

Sahl, Hans-G.; Hahn, Claudia; Brandis, H.

doi:10.1016/s0176-6724(85)80115-2

Cited by 14 publications

(18 citation statements)

References 18 publications

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“…Purified trypsinized cell walls were prepared from S. simulans 22 and M. luteus as previously described (25). Teichoic acids were extracted by hydrolysis with 70% HF for 3 h at 0°C (13).…”

Section: Methodsmentioning

confidence: 99%

“…This has been shown for Pep 5 (25), dihydrostreptomycin (12), and polylysine (data not shown). High ionic strength prevents binding of the peptides to polyanionic compounds (25). As the addition of a sufficient amount of Pep 5 can reverse the binding of the amidase to LTA (2), so mono-and divalent cations, small cationic molecules, and basic peptides are also probably able to prevent binding of the enzyme to or to displace it from teichoic and teichuronic acids, thereby causing activation.…”

Section: Methodsmentioning

confidence: 99%

“…Both were reported to consist of mixtures of closely related, structurally similar peptides (Mr, 3,500; isoelectric point, >10.5) and to contain the rare thioether amino acid lanthionine (10,24). Pep 5 adsorbs to gram-positive bacteria via ionic interaction with negatively charged cell wall polymers (teichoic, teichuronic, and lipoteichoic acids [LTA]) (25). The primary target of Pep 5 and nisin is the cytoplasmic membrane, which becomes permeable for small molecules such as K+, amino acids, and ATP (19,20,23).…”

mentioning

confidence: 99%

“…However, autolysin activity could be demonstrated in the supernatant of a cell suspension of S. simulans treated with Pep 5, including at least two different autolysins which hydrolyzed the glycan chain and peptide bridges of the murein. The strongly basic peptide Pep 5 was shown to associate with the polyanionic LTA (2,25) which have been discussed as potential regulators of autolysin activity (8,18). The autolysins of S. simulans were inhibited in the presence of LTA and were reactivated by addition of a sufficient quantity of Pep 5.…”

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confidence: 99%

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Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase

1987

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Pep 5 and nisin are cationic peptide antibiotics which in addition to their membrane-disruptive action induce autolysis in staphylococci. To investigate the mechanism of lysis induction, the influence of the peptides on the activity of the N-acetylmuramoyl-L-alanine amidase of Staphylococcus simulans 22 was studied. In experiments with isolated cell walls at low ionic strength, the amidase activity was stimulated by the addition of Pep 5 and nisin, as well as by polylysine, streptomycin, and mono-and divalent cations. The concentrations necessary for activation depended on the nature of the cation and rahged from 5 ,uM for poly-L-lysine (n = 17) to 150 mM for Na+ at a cell wail concentration of 100 ,ug of cell wails per ml. No effect was observed if the cell walls were devoid of polyanionic constituents. Kinetic data suggested that the amidase bound to the teichoic and teichuronic acids of the cell wall and was thereby inhibited. Cationic molecules reversed this inhibition, most likely by displacing the enzyme from the polyanions. If the concentrations of the larger peptides were high in relation to cell wall concentration, the activation turned into inhibition, presumably by interfering with the access of the enzyme to its substrate. These experiments demonstrate that the activity of the amidase is modulated by basic peptides in vitro and help to explain how Pep 5 and nisin may cause lysis of treated cells.

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“…Purified trypsinized cell walls were prepared from S. simulans 22 and M. luteus as previously described (25). Teichoic acids were extracted by hydrolysis with 70% HF for 3 h at 0°C (13).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase

1987

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“…This suspension was incorporated into agar plates and used for activity assays. Purified trypsinized cell walls were prepared from M. luteus and S. aureus NMSCC 133 as previously described (21). Teichoic acids were extracted by hydrolysis with 70% HF for 3 h at 0°C (16).…”

Section: Methodsmentioning

confidence: 99%

Purification and Partial Characterization of a Murein Hydrolase, Millericin B, Produced by Streptococcus milleri NMSCC 061

Beukes

Bierbaum

Sahl

et al. 2000

Appl Environ Microbiol

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Streptococcus milleri NMSCC 061 was screened for antimicrobial substances and shown to produce a bacteriolytic cell wall hydrolase, termed millericin B. The enzyme was purified to homogeneity by a four-step purification procedure that consisted of ammonium sulfate precipitation followed by gel filtration, ultrafiltration, and ion-exchange chromatography. The yield following ion-exchange chromatography was 6.4%, with a greater-than-2,000-fold increase in specific activity. The molecular weight of the enzyme was 28,924 as determined by electrospray mass spectrometry. The amino acid sequences of both the N terminus of the enzyme (NH 2 SENDFSLAMVSN) and an internal fragment which was generated by cyanogen bromide cleavage (NH 2 SIQTNAPWGL) were determined by automated Edman degradation. Millericin B displayed a broad spectrum of activity against gram-positive bacteria but was not active against Bacillus subtilis W23 or Escherichia coli ATCC 486 or against the producer strain itself. N-Dinitrophenyl derivatization and hydrazine hydrolysis of free amino and free carboxyl groups liberated from peptidoglycan digested with millericin B followed by thin-layer chromatography showed millericin B to be an endopeptidase with multiple activities. It cleaves the stem peptide at the N terminus of glutamic acid as well as the N terminus of the last residue in the interpeptide cross-link of susceptible strains.Most eubacteria are mechanically stabilized by the shapedetermining peptidoglycan, which consists of polysaccharide chains that are connected by short peptides. Expansion of the bacterial cell wall during growth and splitting of the septum prior to cell separation require enzymes that cleave some of the existing covalent bonds within the peptidoglycan sacculus. These enzymes are collectively known as peptidoglycan hydrolases (24). Some have been characterized as N-acetylmuramidases, N-acetylglucosaminidases, N-acetylmuramyl-L-alanine amidases, endopeptidases, and transglycosidases (10). Most peptidoglycan hydrolases have been characterized as bacteriolytic enzymes by in vitro studies (29). Lysostaphin, which is active against Staphylococcus aureus, is one such enzyme. It has been suggested that lysostaphin may function catabolically to release nutrients from other staphylococci in the environment, with the producer cell being protected by a specific immunity protein (12,18). The lytic effect of lysostaphin results from a direct attack on the integrity of the staphylococcal cell wall, specifically bringing about cleavage of the pentaglycine crosslink between the stem peptides of individual peptidoglycan subunits (4).Zoocin A, a bacteriolytic cell wall hydrolase produced by Streptococcus zooepidemicus 4881, like lysostaphin, cleaves bonds within the peptide moiety of peptidoglycan (26). Zoocin A has been shown to inhibit the growth of all Streptococcus pyogenes strains and all S. zooepidemicus strains other than 4881 itself (14). Both enzymes, lysostaphin and zoocin A, possess a catalytic domain in the N-terminal portion and a...

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Aspergillus and Aspergillosis

Bossche¹,

Mackenzie²,

Cauwenbergh³

1988

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Interaction of the Staphylococcin-like Peptide Pep 5 with Cell Walls and Isolated Cell Wall Components of Gram-Positive Bacteria

Cited by 14 publications

References 18 publications

Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase

Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase

Purification and Partial Characterization of a Murein Hydrolase, Millericin B, Produced by Streptococcus milleri NMSCC 061

Aspergillus and Aspergillosis

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