Previous Article | Next Article 
Journal of Bacteriology, November 1999, p. 6865-6875, Vol. 181, No. 22
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Thickness and Elasticity of Gram-Negative Murein
Sacculi Measured by Atomic Force Microscopy
X.
Yao,1
M.
Jericho,1
D.
Pink,2 and
T.
Beveridge3,*
Department of Physics, Dalhousie University,
Halifax, Nova Scotia, Canada B3H 4J11;
Department of Physics, St. Francis Xavier University,
Antigonish, Nova Scotia, Canada B2G 2W52; and
Department of Microbiology, College of Biological Science,
University of Guelph, Guelph, Ontario, Canada N1G 2W13
Received 28 June 1999/Accepted 7 September 1999
Atomic force microscopy was used to measure the thickness of
air-dried, collapsed murein sacculi from Escherichia coli
K-12 and Pseudomonas aeruginosa PAO1. Air-dried sacculi
from E. coli had a thickness of 3.0 nm, whereas those from
P. aeruginosa were 1.5 nm thick. When rehydrated, the
sacculi of both bacteria swelled to double their anhydrous thickness.
Computer simulation of a section of a model single-layer peptidoglycan
network in an aqueous solution with a Debye shielding length of 0.3 nm
gave a mass distribution full width at half height of 2.4 nm, in
essential agreement with these results. When E. coli
sacculi were suspended over a narrow groove that had been etched into a
silicon surface and the tip of the atomic force microscope used to
depress and stretch the peptidoglycan, an elastic modulus of 2.5 × 107 N/m2 was determined for hydrated
sacculi; they were perfectly elastic, springing back to their original
position when the tip was removed. Dried sacculi were more rigid with a
modulus of 3 × 108 to 4 × 108
N/m2 and at times could be broken by the atomic force
microscope tip. Sacculi aligned over the groove with their long axis at
right angles to the channel axis were more deformable than those with their long axis parallel to the groove axis, as would be expected if
the peptidoglycan strands in the sacculus were oriented at right angles
to the long cell axis of this gram-negative rod. Polar caps were not
found to be more rigid structures but collapsed to the same thickness
as the cylindrical portions of the sacculi. The elasticity of intact
E. coli sacculi is such that, if the peptidoglycan strands
are aligned in unison, the interstrand spacing should increase by 12%
with every 1 atm increase in (turgor) pressure. Assuming an unstressed
hydrated interstrand spacing of 1.3 nm (R. E. Burge, A. G. Fowler, and D. A. Reaveley, J. Mol. Biol. 117:927-953, 1977) and
an internal turgor pressure of 3 to 5 atm (or 304 to 507 kPa) (A. L. Koch, Adv. Microbial Physiol. 24:301-366, 1983), the natural
interstrand spacing in cells would be 1.6 to 2.0 nm. Clearly, if large
macromolecules of a diameter greater than these spacings are secreted
through this layer, the local ordering of the peptidoglycan must
somehow be disrupted.
*
Corresponding author. Mailing address: Department of
Microbiology, College of Biological Science, University of Guelph,
Guelph, Ontario, Canada N1G 2W1. Phone: (519) 824-4120, ext. 3366. Fax: (519) 837-1802. E-mail: tjb{at}micro.uoguelph.ca.
Journal of Bacteriology, November 1999, p. 6865-6875, Vol. 181, No. 22
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Vadillo-Rodriguez, V., Schooling, S. R., Dutcher, J. R.
(2009). In Situ Characterization of Differences in the Viscoelastic Response of Individual Gram-Negative and Gram-Positive Bacterial Cells. J. Bacteriol.
191: 5518-5525
[Abstract]
[Full Text]
-
Mannik, J., Driessen, R., Galajda, P., Keymer, J. E., Dekker, C.
(2009). From the Cover: Bacterial growth and motility in sub-micron constrictions. Proc. Natl. Acad. Sci. USA
106: 14861-14866
[Abstract]
[Full Text]
-
Bergholz, P. W., Bakermans, C., Tiedje, J. M.
(2009). Psychrobacter arcticus 273-4 Uses Resource Efficiency and Molecular Motion Adaptations for Subzero Temperature Growth. J. Bacteriol.
191: 2340-2352
[Abstract]
[Full Text]
-
Huang, K. C., Mukhopadhyay, R., Wen, B., Gitai, Z., Wingreen, N. S.
(2008). Cell shape and cell-wall organization in Gram-negative bacteria. Proc. Natl. Acad. Sci. USA
105: 19282-19287
[Abstract]
[Full Text]
-
Gan, L., Chen, S., Jensen, G. J.
(2008). Molecular organization of Gram-negative peptidoglycan. Proc. Natl. Acad. Sci. USA
105: 18953-18957
[Abstract]
[Full Text]
-
Stukalov, O., Korenevsky, A., Beveridge, T. J., Dutcher, J. R.
(2008). Use of Atomic Force Microscopy and Transmission Electron Microscopy for Correlative Studies of Bacterial Capsules. Appl. Environ. Microbiol.
74: 5457-5465
[Abstract]
[Full Text]
-
Vadillo-Rodriguez, V., Beveridge, T. J., Dutcher, J. R.
(2008). Surface Viscoelasticity of Individual Gram-Negative Bacterial Cells Measured Using Atomic Force Microscopy. J. Bacteriol.
190: 4225-4232
[Abstract]
[Full Text]
-
Borgnia, M. J., Subramaniam, S., Milne, J. L. S.
(2008). Three-Dimensional Imaging of the Highly Bent Architecture of Bdellovibrio bacteriovorus by Using Cryo-Electron Tomography. J. Bacteriol.
190: 2588-2596
[Abstract]
[Full Text]
-
Lan, G., Wolgemuth, C. W., Sun, S. X.
(2007). Z-ring force and cell shape during division in rod-like bacteria. Proc. Natl. Acad. Sci. USA
104: 16110-16115
[Abstract]
[Full Text]
-
Chen, I., Christie, P. J., Dubnau, D.
(2005). The Ins and Outs of DNA Transfer in Bacteria. Science
310: 1456-1460
[Abstract]
[Full Text]
-
Zahrl, D., Wagner, M., Bischof, K., Bayer, M., Zavecz, B., Beranek, A., Ruckenstuhl, C., Zarfel, G. E., Koraimann, G.
(2005). Peptidoglycan degradation by specialized lytic transglycosylases associated with type III and type IV secretion systems. Microbiology
151: 3455-3467
[Abstract]
[Full Text]
-
Ma, H., Snook, L. A., Kaminskyj, S. G. W., Dahms, T. E. S.
(2005). Surface ultrastructure and elasticity in growing tips and mature regions of Aspergillus hyphae describe wall maturation. Microbiology
151: 3679-3688
[Abstract]
[Full Text]
-
Gaboriaud, F., Bailet, S., Dague, E., Jorand, F.
(2005). Surface Structure and Nanomechanical Properties of Shewanella putrefaciens Bacteria at Two pH values (4 and 10) Determined by Atomic Force Microscopy. J. Bacteriol.
187: 3864-3868
[Abstract]
[Full Text]
-
Zhao, L., Schaefer, D., Marten, M. R.
(2005). Assessment of Elasticity and Topography of Aspergillus nidulans Spores via Atomic Force Microscopy. Appl. Environ. Microbiol.
71: 955-960
[Abstract]
[Full Text]
-
Li, G., Smith, C. S., Brun, Y. V., Tang, J. X.
(2005). The Elastic Properties of the Caulobacter crescentus Adhesive Holdfast Are Dependent on Oligomers of N-Acetylglucosamine. J. Bacteriol.
187: 257-265
[Abstract]
[Full Text]
-
Vollmer, W., Holtje, J.-V.
(2004). The Architecture of the Murein (Peptidoglycan) in Gram-Negative Bacteria: Vertical Scaffold or Horizontal Layer(s)?. J. Bacteriol.
186: 5978-5987
[Full Text]
-
Dufrene, Y. F.
(2004). Refining Our Perception of Bacterial Surfaces with the Atomic Force Microscope. J. Bacteriol.
186: 3283-3285
[Full Text]
-
Matias, V. R. F., Al-Amoudi, A., Dubochet, J., Beveridge, T. J.
(2003). Cryo-Transmission Electron Microscopy of Frozen-Hydrated Sections of Escherichia coli and Pseudomonas aeruginosa. J. Bacteriol.
185: 6112-6118
[Abstract]
[Full Text]
-
Touhami, A., Hoffmann, B., Vasella, A., Denis, F. A., Dufrene, Y. F.
(2003). Aggregation of yeast cells: direct measurement of discrete lectin-carbohydrate interactions. Microbiology
149: 2873-2878
[Abstract]
[Full Text]
-
Dmitriev, B. A., Toukach, F. V., Schaper, K.-J., Holst, O., Rietschel, E. T., Ehlers, S.
(2003). Tertiary Structure of Bacterial Murein: the Scaffold Model. J. Bacteriol.
185: 3458-3468
[Abstract]
[Full Text]
-
Dufrene, Y. F.
(2002). Atomic Force Microscopy, a Powerful Tool in Microbiology. J. Bacteriol.
184: 5205-5213
[Full Text]
-
Ward, D. V., Draper, O., Zupan, J. R., Zambryski, P. C.
(2002). Inaugural Article: Peptide linkage mapping of the Agrobacterium tumefaciens vir-encoded type IV secretion system reveals protein subassemblies. Proc. Natl. Acad. Sci. USA
99: 11493-11500
[Abstract]
[Full Text]
-
Glasauer, S., Langley, S., Beveridge, T. J.
(2001). Sorption of Fe (Hydr)Oxides to the Surface of Shewanella putrefaciens: Cell-Bound Fine-Grained Minerals Are Not Always Formed De Novo. Appl. Environ. Microbiol.
67: 5544-5550
[Abstract]
[Full Text]
-
Bayer, M., Iberer, R., Bischof, K., Rassi, E., Stabentheiner, E., Zellnig, G., Koraimann, G.
(2001). Functional and Mutational Analysis of P19, a DNA Transfer Protein with Muramidase Activity. J. Bacteriol.
183: 3176-3183
[Abstract]
[Full Text]
-
Pink, D., Moeller, J., Quinn, B., Jericho, M., Beveridge, T.
(2000). On the Architecture of the Gram-Negative Bacterial Murein Sacculus. J. Bacteriol.
182: 5925-5930
[Abstract]
[Full Text]