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Freeze-etching of the outer membranes of Pseudomonas and Acinetobacter

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Abstract

The location and surface structure of the fracture faces revealed by freeze-etching within the cell walls of Pseudomonas fluorescens NCTC 10038 and Acinetobacter sp. strain MJT/F5/5 have been compared.

Treatment of intact cells with lysozyme and ethylenediaminetetraacetic acid, followed by stabilization with MgCl2, detaches the outer membranes of both organisms from underlying layers of the envelope. In the Acinetobacter, the detached outer membrane (om) retains the a-layer of hexagonally-arranged subunits on its outer surface, whereas the Pseudomonas has no a-layer.

Freeze-etching of these detached structures reveals the etched outer and inner surfaces of the outer membranes (in the Pseudomonas) or the om + a-layers (in the Acinetobacter). In addition, the detached structures in both organisms fractured along an internal plane.

Since the structures of these internal fracture faces are identical with those of fracture faces produced within the cell walls of intact cells, freeze-etched in the presence of glycerol, it is clear that, under these conditions also, the main fracture plane occurs within the outer membranes.

The technique used also reveals the etched outer surface of the plasma membranes, as well as the internal fracture faces which are normally seen, and these are illustrated.

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Authors and Affiliations

  1. Strangeways Research Laboratory, Cambridge

    Margaret J. Thornley & U. B. Sleytr

  2. Immunology Division Dept. of Pathology, Addenbrookes Hospital, Hills Rd., Cambridge, England

    Margaret J. Thornley

Authors
  1. Margaret J. Thornley
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  2. U. B. Sleytr
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Thornley, M.J., Sleytr, U.B. Freeze-etching of the outer membranes of Pseudomonas and Acinetobacter . Arch. Microbiol. 100, 409–417 (1974). https://doi.org/10.1007/BF00446332

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  • DOI: https://doi.org/10.1007/BF00446332

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