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Journal of Bacteriology, November 2008, p. 7414-7418, Vol. 190, No. 22
0021-9193/08/$08.00+0     doi:10.1128/JB.00581-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Lipoteichoic Acid Is a Major Component of the Bacillus subtilis Periplasm{triangledown}

Valério R. F. Matias* and Terry J. Beveridge

Department of Molecular and Cellular Biology, College of Biological Science, and Advanced Foods and Materials Network—Network Centres of Excellence (AFMnet-NCE), University of Guelph, Guelph, Ontario N1G 2W1, Canada

Received 27 April 2008/ Accepted 3 September 2008

Cryo-electron microscopy (cryo-EM) of frozen-hydrated specimens allows high-resolution observation of structures in optimally preserved samples. In gram-positive bacteria, this method reveals the presence of a periplasmic space between the plasma membrane and an often differentiated cell wall matrix. Since virtually nothing is known about the composition of its constituent matter (i.e., the periplasm), it is still unclear what structures (or mechanism) sustain a gram-positive periplasmic space. Here we have used cryo-EM of frozen-hydrated sections in combination with various labels to probe the model gram-positive organism Bacillus subtilis for major periplasmic components. Incubation of cells with positively charged gold nanoparticles showed almost similar levels of gold binding to the periplasm and the cell wall. On cells whose cell walls were enzymatically hydrolyzed (i.e., on protoplasts), a surface diffuse layer extending ~30 nm from the membrane was revealed. The thickness and density of this layer were not significantly altered after treatment with a nonspecific protease, whereas it was labeled with anti-lipoteichoic acid (LTA) antibodies conjugated to nanogold. Further, the LTA layer spans most of the thickness of the periplasmic space, which strongly suggests that LTA is a major component of the B. subtilis periplasm.

* Corresponding author. Present address: Max Planck Institute of Biochemistry, Department of Molecular Structural Biology, Am Klopferspitz 18, D-82152, Martinsried, Germany. Phone: 49 (89) 8578-2032. Fax: 49 (89) 8578-3777. E-mail: matias{at}biochem.mpg.de

{triangledown} Published ahead of print on 12 September 2008.

Journal of Bacteriology, November 2008, p. 7414-7418, Vol. 190, No. 22
0021-9193/08/$08.00+0     doi:10.1128/JB.00581-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Rahman, O., Pfitzenmaier, M., Pester, O., Morath, S., Cummings, S. P., Hartung, T., Sutcliffe, I. C. (2009). Macroamphiphilic Components of Thermophilic Actinomycetes: Identification of Lipoteichoic Acid in Thermobifida fusca. J. Bacteriol. 191: 152-160 [Abstract] [Full Text]  


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