Time-Lapse Cinematography of Vancomycin-Treated Microbial Cells

Durham, Norman N.; Noller, Eric C.; Burger, Mike W.; Best, Gary K.

doi:10.1139/m67-054

Cited by 6 publications

(3 citation statements)

References 8 publications

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“…the cell wall. The spectral changes observed in incomycin the 15N NMR spectra of vancomycin-intoxicated tracted by B. licheniformis resemble those associated with cell walls plasmolysis and support the suggestions of Durwalls and ham et al (9) and Marquis (22 …”

supporting

confidence: 84%

Effects of Binding and Bactericidal Action of Vancomycin on Bacillus licheniformis Cell Wall Organization as Probed by ¹⁵ N Nuclear Magnetic Resonance Spectroscopy

Irving

Lapidot

1978

Antimicrob Agents Chemother

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The effects of binding and the bactericidal action of vancomycin on the arrangement and mobilities of cell wall polymers in Bacillus licheniformis were investigated by '6N nuclear magnetic resonance spectroscopy. The bactericidal action of vancomycin led to reduced mobilities of cell wall teichoic acid and teichuronic acid in surviving cells. The decrease in teichoic acid mobility was also observed upon binding of vancomycin to B. licheniformis cells and resulted from a specific interaction between the antibiotic and teichoic acid, rather than from electrostatic contraction of the cell wall. The reduction in teichuronic acid mobility appeared to be related either to the elastic contraction of the cell wall resulting from loss of cell turgor or to separation of the cell wall from the protoplast membrane. No spectral changes associated with cell wall autolysis or alterations in cell wall composition, amidation, and cross-linking were found in vancomycin-treated B. licheniformis cells. Binding of vancomycin to Micrococcus lysodeikticus cell walls led to a decrease in mobility of C-terminal D-alanine residues but was accompanied by an increase in the mobilities of other peptidoglycan residues. The possible contributions of changes in the arrangements of cell wall polymers to the lethal action of vancomycin is discussed.The bactericidal actions of many antibiotics are associated with the inhibitions of cell wall biosynthesis that result from the binding of the antibiotics to enzymes and intermediates along the cell wall biosynthetic pathway (32). Rogers and a co-worker have suggested (28, 29) that when cell wall biosynthesis is inhibited, the natural ongoing autolysis of the cell walltis no longer in balance with the insertion of new cell material. Hydrolytic cleavage of peptidoglycan glycosidic and peptide bonds proceeds until the mechanical properties of the cell wall that are responsible for osmotic protection of the protoplast membrane are lost and the membrane is damaged.The role of autolysins in the actions of penicillin and vancomycin has been amply demonstrated by the loss of the lethal actions of the antibiotics when cell wall autolysins are inhibited (1,28,29,32,34). Vancomycin forms strong complexes with gram-positive bacterial cell walls and lipidsoluble cell wall intermediates and peptidoglycan precursors in the protoplast membrane (13,18,23,24,26,37). The inhibition of peptidoglycan biosynthesis by vancomycin in cell-free systems is restricted to its complexing with lipidsoluble intermediates in the membrane fraction (12,13,18). However, in the whole cell, part of the bactericidal effect of vancomycin may also be associated with the strong binding of vancomycin to the cell wall (2, 3, 13). The binding of highly charged vancomycin molecules (2, 3) to the amphoteric polyelectrolyte gel-like cell wall (22) is likely to result in changes in the threedimensional arrangements of the cell wall polymers and in the contacts between the cell wall and protoplast membrane, which could affect the relative rates of c...

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supporting

confidence: 84%

Effects of Binding and Bactericidal Action of Vancomycin on Bacillus licheniformis Cell Wall Organization as Probed by ¹⁵ N Nuclear Magnetic Resonance Spectroscopy

Irving

Lapidot

1978

Antimicrob Agents Chemother

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“…Durham et al (8) microscopically observed shrinkage of whole B. cereus cells exposed to vancomycin. They considered that this shrinkage was due to elastic contraction of the wall after loss of cell turgor.…”

Section: Resultsmentioning

confidence: 99%

Salt-induced Contraction of Bacterial Cell Walls

Marquis

1968

J Bacteriol

129

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Intact Bacillus megaterium cells were found to contract as much as 26% in terms of dextran-impermeable volume when transferred from water to unbuffered, nonplasmolyzing NaCl solutions. This shrinkage appeared to be primarily due to electrostatic wall contraction rather than to any osmotic response of the cells. A variety of salts (but not sucrose) added to water suspensions of isolated cell walls caused protons to be released from the walls with resultant lowering of suspension pH and contraction of the structures. In effect, B. megaterium walls behaved as flexible, amphoteric polyelectrolytes, and their compactness in aqueous suspensions was affected by changes in environmental ionic strength and pH. Isolated walls were most compact in low ionic strength media with a pH of about 4, a value close to the apparent isoelectric pH of wall peptidoglycan. Electrostatic attractions appeared to play a major role in determining the compactness of highly contracted walls, and the walls responded to increased environmental ionic strength by expanding. In contrast, electrostatic repulsions were dominant in highly expanded walls, and increased environmental ionic strength induced wall contraction. Walls of whole bacteria also shrank when the cells were plasmolyzed. This second type of contraction seemed to result from relief of wall tension during plasmolysis, and it could be induced with nonionic solutes. Thus, cell wall tone in B. megaterium appeared to be set both by mechanical tension and by electrostatic interactions among wall ions.Many investigations have shown shrinkage of whole bacterial cells after transfer from dilute to concentrated salt solutions. For example, Johnson and Harvey (13) concluded from measurements of packed cell volume and light scattering that gram-negative Achromobacter (Photobacterium) fischeri cells shrank as much as 27% in volume when transferred from seawater to 2.5 times concentrated seawater. Salt-induced contraction of many other gram-negative bacteria has been reported (3,4,12,15), and there appeared to be a close association between whole cell shrinkage and observable plasmolysis.Gram-positive bacteria also can be made to shrink in salt solutions, even though they are not readily plasmolyzed. Dutky (unpublished data, cited in reference 14) found that Bacillus megaterium cells contracted from 15 to 25% when transferred from normal broth to broth containing 2 M NaCl. Mitchell and Moyle (18) indicated that Staphylococcus aureus cells contracted from 2 to 3% in linear dimensions, without becoming visibly plasmolyzed, when transferred from 0.1 to 1.0 M NaCl solution.Salt-induced shrinkage of bacteria has been interpreted as an osmotic response of the cells mainly because the effect can be produced in easily plasmolyzed, gram-negative organisms with either electrolytes or nonelectrolytes (3,4,13,15). Our results, however, indicate that salt-induced contraction of gram-positive B. megaterium cells does not primarily involve osmotic plasmolysis, but that it is caused mainly by electrostatic contr...

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“…Although TLM is mostly used with cultured mammalian cells and live cells in tissues, the significant number of reports indicates that TLM could be employed to observe and study prokaryotic cells and other unicellular and multicellular organisms as well as viruses. Here, we mention only few examples, such as time-lapse imaging of growth, cell-cell contacts and formation of spherical granules in E. coli [191][192][193][194]; time-lapse visualization of bacterial colony morphologies in the special bacterial chamber MOCHA [195]; screening and assessing effects of antibiotics, such as antibiotics-bacteria interactions [196][197][198][199] and studying yeasts [200][201][202] and viruses [203][204][205][206][207]. The smaller microorganisms, analogously to intracellular structures, usually require higher magnification and more sophisticated microscopic equipment.…”

Section: Time-lapse Microscopy: From Making Movies To Bedside 21 Vementioning

confidence: 99%

Time-Lapse Microscopy

Collins¹,

Knippenberg²,

Ding³

et al. 2019

Cell Culture

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Time-Lapse Cinematography of Vancomycin-Treated Microbial Cells

Cited by 6 publications

References 8 publications

Effects of Binding and Bactericidal Action of Vancomycin on Bacillus licheniformis Cell Wall Organization as Probed by ¹⁵ N Nuclear Magnetic Resonance Spectroscopy

Effects of Binding and Bactericidal Action of Vancomycin on Bacillus licheniformis Cell Wall Organization as Probed by ¹⁵ N Nuclear Magnetic Resonance Spectroscopy

Salt-induced Contraction of Bacterial Cell Walls

Time-Lapse Microscopy

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Time-Lapse Cinematography of Vancomycin-Treated Microbial Cells

Cited by 6 publications

References 8 publications

Effects of Binding and Bactericidal Action of Vancomycin on Bacillus licheniformis Cell Wall Organization as Probed by 15 N Nuclear Magnetic Resonance Spectroscopy

Effects of Binding and Bactericidal Action of Vancomycin on Bacillus licheniformis Cell Wall Organization as Probed by 15 N Nuclear Magnetic Resonance Spectroscopy

Salt-induced Contraction of Bacterial Cell Walls

Time-Lapse Microscopy

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Product

Resources

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Effects of Binding and Bactericidal Action of Vancomycin on Bacillus licheniformis Cell Wall Organization as Probed by ¹⁵ N Nuclear Magnetic Resonance Spectroscopy

Effects of Binding and Bactericidal Action of Vancomycin on Bacillus licheniformis Cell Wall Organization as Probed by ¹⁵ N Nuclear Magnetic Resonance Spectroscopy