Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase.
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 ranged from 5 microM for poly-L-lysine (n = 17) to 150 mM for Na+ at a cell wall concentration of 100 micrograms of cell walls 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.
D000581:Amidohydrolases / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001430:Bacteriocins; D001433:Bacteriolysis; D002473:Cell Wall / Q000201:enzymology / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000378:metabolism*; D009561:Nisin / Q000494:pharmacology*; D010455:Peptides / Q000494:pharmacology; D011107:Polylysine / Q000494:pharmacology; D013210:Staphylococcus / Q000201:enzymology* / Q000378:metabolism; D013682:Teichoic Acids / Q000378:metabolism; D014574:Uronic Acids / Q000378:metabolism
D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D010455:Peptides; D013682:Teichoic Acids; D014574:Uronic Acids; D009561:Nisin; D011107:Polylysine; C010788:teichuronic acid; C054222:lantibiotic Pep5; D000581:Amidohydrolases; D009238:N-Acetylmuramoyl-L-alanine Amidase