Abstract
To analyze if chemical cell wall alterations contribute to penicillin-induced bacteriolysis, changes in the amount, stability, and chemical composition of staphylococcal cell walls were investigated. All analyses were performed before onset of bacteriolysis i.e. during the first 60 min following addition of different penicillin G doses. Only a slight reduction of the amount of cell wall material incorporated after penicillin addition at the optimal lytic concentration was observed as compared to control cells. However, the presence of higher penicillin G concentrations reduced the incorporation of wall material progressively without bacteriolysis. Losses of wall material during isolation of dodecylsulfate insoluble cell walls were monitored to assess the stability of the wall material following penicillin addition. Wall material grown at the lytic penicillin concentration was least stable but about 30% of the newly incorporated wall material withstood even the harsh conditions of mechanical breakage and dodecylsulfate treatment. Dodecylsulfate insoluble cell walls were used for chemical analyses. While peptidoglycan chain length was unaffected in the presence of penicillin, other wall parameters were considerably altered: peptide cross-linking was reduced in the wall material synthesized after addition of penicillin; reductions from approx. 85% in controls to about 60% were similar for lytic and also for very high penicillin concentrations leading to nonlytic death. O-acetylation was also reduced after treatment with penicillin; this effect paralleled the occurence of subsequent bacteriolysis at different drug concentrations. The results are not consistent with hypotheses explaining penicillin-induced lysis as a result of an overall weakened cell wall structure or an overall activation of autolytic wall enzymes but not conflicting with the model that ascribes penicillin-induced bacteriolysis as the result of a very restricted, local perforation of the peripheral cell wall (murosome-induced bacteriolysis).
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Abbreviations
- CL:
-
Cross-linking
- DNFB:
-
2,4-dinitro-1-fluorobenzole
- MIC:
-
Minimal inhibitory concentration
- OD:
-
Optical density at 578 nm
- PEN:
-
Penicillin G
References
Andrew AL; Perkins HR (1987) Progress of O-acetylation and cross-linking of peptidoglycan in Neisseria gonorrhoeae grown in the presence of penicillin. Gen Microbiol 133:1743–1750
Beise F, Labischinski H, Giesbrecht P (1988a) Role of Penicillin-binding proteins of Staphylococcus aureus in the induction of bacteriolysis by β-lactam antibiotics. In: Actor P, Daneo-Moore L, Higgins ML, Salton MRJ, Shockman GD (eds) Antibiotic inhibition of bacterial cell surface assembly and function. American Society for Microbiology, pp 360–366
Beise F, Labischinski H, Giesbrecht P (1988b) Selective inhibition of Penicillin-binding proteins and its effects on growth and architecture of Staphylococcus aureus. FEMS Microbiol Lett 55:195–202
Blümel P, Uecker W, Giesbrecht P (1981) In vitro studies on the possible role of cell wall turnover in Staphylococcus aureus during infection. In: Jeliaszewicz J (ed) Staphylococci and staphylococcal infections. Zbl Bakt Suppl 10 Gustav Fischer, Stuttgart, pp 435–439
Blundell JK, Perkins HR (1981) Effects of β-lactam antibiotics on peptidoglycan synthesis in growing Neisseria gonorrhoeae, including changes in the degree of O-acetylation. J Bacteriol 147:633–641
Blundell JK, Perkins HR (1986) Selectivity for O-acetylated peptidoglycan during endopeptidase action by permeabilized Neisseria gonorrhoeae EFMS Microbiol Lett 30:67–69
Dezellee P, Shockman GD (1975) Studies of the formation of peptide crosslinks in the cell wall peptidoglycan of Streptococcus faecalis. J Biol Chem 250:6806–6810
Eagle H, Musselman AD (1948) The rate of bactericidal action of penicillin in vitro as a function of its concentration, and its paradoxically reduced activity at high concentrations against certain organisms. J Exp Med 88:99–131
Fischer H, Tomasz A (1984) Peptidoglycan cross-linking and teichoic acid attachment in Streptococcus pneumoniae. J Bacteriol 163:46–54
Fordham WD, Gilvarg C (1974) Kinetics of cross-linking of peptidoglycan in Bacillus megaterium. J Biol Chem 249:2478–2483
Fromme I, Beilharz H (1978) Gas chromatographic assay of total and O-acetyl groups in bacterial lipopolysaccharides. Anal Biochem 84:347–353
Ghuysen JM, Tipper J, Strominger JL (1966) Enzymes that degrade bacterial cell walls. Methods Enzymol 8:685–699
Giesbrecht P, Morioka H, Krüger D, Kersten T, Wecke J (1983) Restoration of penicillin-damaged cell walls by de novo murein synthesis and successive murein degradation in staphylococci, revealing a hitherto unknown mechanism of penicillin action: blockage of autolytic wall processes by penicillin. In: Hakenbeck R, Höltje JV, Labischinski H (eds) The target of penicillin The murein sacculus of bacterial cell walls. Architecture and growth. De Gruyter, Berlin New York, pp 243–248
Giesbrecht P (1984) Novel bacterial organelles (“murosomes”) in staphylococci: their involvement in wall assembly. In: Nombela C (ed) Microbial cell wall synthesis and autolysis. Elsevier Science Publ, Amsterdam pp 177–186
Giesbrecht P, Labischinski H, Wecke J (1985) A special morphogenetic wall defect and the subsequent activity of “murosomes” as the very reason for penicillin-induced bacteriolysis in staphylococci. Arch Microbiol 141:315–324
Gmeiner J, Kroll HP (1981) Murein biosynthesis and O-acetylation of N-acetylmuramic acid during the cell division cycle of Proteus mirabilis Eur J Biochem 117:171–177
Johannsen L (1982) Über den Einfluß der O-Acetylierung des Mureins von Staphylococcus aureus auf die Abbaubarkeit bakterieller Zellwände durch zellwandlytische Enzymsysteme. Thesis, FU Berlin
Johannsen L, Labischinski H, Reinicke B, Giesbrecht P (1983) Changes in the chemical structure of walls of Staphylococcus aureus grown in the presence of chloramphenicol. FEMS Microbiol Lett 16:313–316
Labischinski H, Maidhof H, Franz M, Krüger D, Sidow T, Giesbrecht P (1988) Biochemical and biophysical investigations into the cause of penicillin-induced lytic death of staphylococci: checking the predictions of the murosome model In: Actor P, Daneo-Moore L, Higgins L, Salton MRJ, Shockman GD (eds) Antibiotic inhibition of bacterial cell surface assembly and function. American Society for Microbiology, pp 242–257
Lear AL, Perkins HR (1987) Progress of O-acetylation and cross-linking of peptidoglycan in Neisseria gonorrhoeae grown in the presence of penicillin. J Gen Microbiol 133:1743–1750
Maidhof H, Johannsen L, Labischinski H, Giesbrecht P (1989) Onset of penicillin-induced bacteriolysis in staphylococci is cell cycle dependent. J Bacteriol 171:2252–2257
Martin H, Gmeiner J (1979) Modification of peptidoglycan structure by penicillin action in cell walls of Proteus mirabilis. Eur J Biochem 95:487–495
Reinicke B, Blümel P, Labischinski H, Giesbrecht P (1985) Neither an enhancement of total autolytic wall degradation nor an inhibition of the incorporation of cell wall material are pre-requisites for penicillin-induced bacteriolysis in staphylococci. Arch Microbiol 141:309–314
Reynolds PE (1988) The essential nature of staphylococcal penicillin-binding proteins. In: Actor P, Daneo-Moore L, Higgins ML, Salton MRJ, Shockman GD (eds) Antibiotic inhibition of bacterial cell surface assembly and function. American Society for Microbiology, pp 334–351
Schaefer F, Christenson JG, Talbot MK (1989) Peptidoglycan synthesis in ether-treated cells of Staphylococcus aureus. Abstract No. K 165. Abstracts of the 1989 Annual Meeting of the American Society of Microbiology, p 272
Schleifer KH, Kandler O (1972) Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriol Rev 36:407–477
Shockman GD, Daneo-Moore L, McDowell TD, Wong W (1981) Function and structure of the cell wall-its importance in the life and death of bacteria. In: Salton M, Shockman GD (eds) β-lactam antibiotics. Mode of action, new developments, and future prospects. Academic Press, New York, pp 31–65
Tipper DJ, Strominger JL (1965) Mechanism of action of penicillins: A proposal based on their structural similarity to acyl-d-alanyl-d-alanine. Proc Natl Acad Sci USA 54:1113–1141
Tipper DJ, Strominger JL (1968) Biosynthesis of the peptidoglycan of bacterial cell walls. J Biol Chem 243:3169–3179
Tomasz A (1979) The mechanism of the irreversible antimicrobial effects of penicillin: how β-lactam antibiotica kill and lyse bacteria. Annu Rev Microbiol 33:113–137
Wecke J, Giesbrecht P (1981) Electron microscopic studies on the “paradoxial” reaction of staphylococci during treatment with antibiotics. In: Jeljaszewicz J (ed) Staphylococci and staphylococcal infections. Zbl Bakt Suppl 10. Gustav Fischer, Stuttgart, pp 435–439
Weidel W, Pelzer H (1964) Bagshaped macromolecules — a new outlock on bacterial cell walls. Adv Enzymol 26:193–232
Wyke A, Ward JB, Hayes MV, Curtis NAC (1981) A role in vivo for penicillin-binding-protein-4 of Staphylococcus aureus. Eur J Biochem 119:389–393
Qoronfleh MW, Wilkinson BJ (1986) Effects of growth of methicillin-resistant and-susceptible Staphylococcus aureus in the presence of β-lactams on peptidoglycan structure and susceptibility to lytic enzymes. Antimicrob Agents Chemother 29:250–257
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Sidow, T., Johannsen, L. & Labischinski, H. Penicillin-induced changes in the cell wall composition of Staphylococcus aureus before the onset of bacteriolysis. Arch. Microbiol. 154, 73–81 (1990). https://doi.org/10.1007/BF00249181
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DOI: https://doi.org/10.1007/BF00249181