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J Biol Chem, Vol. 273, Issue 25, 15661-15666, June 19, 1998

Evidence for a New Type of Outer Membrane Lipid in Oral Spirochete Treponema denticola
FUNCTIONING PERMEATION BARRIER WITHOUT LIPOPOLYSACCHARIDES

Christian P. Schultz, Violetta Wolf^¶, Robert Lange^¶, Erich Mertens, Jörg Wecke, Dieter Naumann, and Ulrich Zähringer

From the  Robert Koch-Institut, D-13353 Berlin, Nordufer 20, ^¶ Immanuel Krankenhaus, D-14109 Berlin, Koenigstrasse 63, and  Forschungszentrum Borstel, Zentrum für Medizin und Biowissenschaften, D-23845 Borstel, Parkallee 1-40, Germany

A new class of outer membrane lipid (OML) was isolated from the oral spirochete Treponema denticola strain ATCC 33521 using a phenol/chloroform/light petroleum procedure normally applied for lipopolysaccharide extraction. In addition to chemical analysis, Fourier transform infrared (FTIR) spectroscopy was applied to compare the biophysical properties of OML with lipopolysaccharides (LPS) and lipoteichoic acids (LTA). Isolated OML fractions represent 1.4% of the total dry cell weight, are about 4 kDa in size, and contain 6% amino sugars, 8% neutral sugars, 14% phosphate, 35% carbazol-positive compounds, and 11% fatty acids (containing iso- and anteiso-fatty acyl chains). Rare for outer membrane lipids, OML contains no significant amount of 3-deoxy-D-manno-octulosonic acids, heptoses, and -hydroxy fatty acids. The fatty acyl chain composition, being similar to that of the cytoplasmic membrane, is quite heterogeneous with anteiso-pentadecanoic acid (12%), palmitic acid (51%), and iso-palmitic acid (19%) as the predominant fatty acids present. Findings of a glycerol-hexose unit and two glycerol-hexadecanoic acid fragments indicate a glycolipid membrane anchor typically found in LTA. There was also no evidence for the presence of a sphingosine-based lipid structure. The results of FTIR measurements strongly suggest that the reconstituted lipid forms normal bilayer structures (vesicles) expressing a high membrane state of order with a distinct phase transition as typical for isolated LPS. However, in contrast to LPS, OML of T. denticola has a lower T_m near 22 °C and a lower cooperativity of the phase transition. The results suggest a different kind of permeation barrier that is built up by this particular OML of T. denticola, which is quite different from LPS normally essential for Gram-negative bacteria.

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Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.

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