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J Biol Chem, Vol. 274, Issue 24, 16819-16824, June 11, 1999
,, and
From the Divisions of The lipopolysaccharide (LPS) of Chlamydia
trachomatis L2 was isolated from tissue culture-grown elementary
bodies using a modified phenol/water procedure followed by extraction
with phenol/chloroform/light petroleum. From a total of 5 × 104 cm2 of infected monolayers, 22.3 mg of LPS
were obtained. Compositional analysis indicated the presence of
3-deoxy-D-manno-oct-2-ulopyranosonic acid
(Kdo), GlcN, phosphorus, and fatty acids in a molar ratio of
2.8:2:2.1:4.5. Matrix-assisted laser-desorption ionization mass
spectrometry performed on the de-O-acylated LPS gave a
major molecular ion peak at m/z 1781.1 corresponding to a molecule of 3 Kdo, 2 GlcN, 2 phosphates, and two
3-hydroxyeicosanoic acid residues. The structure of deacylated LPS
obtained after successive treatment with hydrazine and potassium
hydroxide was determined by 600 MHz NMR spectroscopy as
Kdo Medical and Biochemical
Microbiology and § Biophysics, Research Center Borstel,
Center for Medicine and Biosciences, D-23845 Borstel, Germany
2
8Kdo24Kdo26D-GlcpN
16D-GlcpN 1,4'-bisphosphate. These data, together with those published recently on the acylation pattern of chlamydial lipid A (Qureshi, N., Kaltashov, I., Walker, K., Doroshenko, V., Cotter, R. J., Takayama, K,
Sievert, T. R., Rice, P. A., Lin, J.-S. L., and
Golenbock, D. T. (1997) J. Biol. Chem. 272, 10594-10600) allow us to present for the first time the complete
structure of a major molecular species of a chlamydial LPS.
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