The lipid A from Comamonas testosteroni has been isolated and its complete chemical structure determined [Iida, T., Haishima, Y., Tanaka, A., Nishijima, K., Saito, S. & Tanamoto, K. (1996). Eur J Biochem 237, 468–475]. In this work, the relationship between its chemical structure and biological activity was studied. The lipid A was highly homogeneous chemically and was characterized by the relatively short chain length (C10) of the 3-hydroxy fatty acid components directly bound to the glucosamine disaccharide backbone by either amide or ester linkages. The lipid A exhibited endotoxic activity in all of the assay systems tested (mitogenicity in mouse spleen cells; induction of tumour necrosis factor alpha release from both mouse peritoneal macrophages and mouse macrophage-like cell line J774-1, as well as from the human monocytic cell line THP-1; induction of nitric oxide release from J774-1 cells; Limulus gelation activity and lethal toxicity in galactosamine-sensitized mice) to the same extent as did ‘Salmonella minnesota’ lipid A or Escherichia coli LPS used as controls. The strong endotoxic activity of the C. testosteroni lipid A indicates that the composition of 3-hydroxydecanoic acid is not responsible for the low endotoxicity of the lipid A observed in members of the genus Rhodopseudomonas, as has previously been suggested. Furthermore, both the lack of a second acylation of the 3-hydroxy fatty acid attached at the 3′ position, and the substitution of the hydroxyl group of the 3-hydroxy fatty acid attached at position 2, do not affect the manifestation of endotoxic activity or species specificity.
GalanosC.,
WestphalO,
LüderitzO.1971; Preparations and properties of antisera against the lipid A component of bacterial lipopolysaccharides. Eur J Biochem 24:116–122[CrossRef]
GalanosC.,
FreudenbergM. A.,
ReutterW.
1979; Galactosamine-induced sensitization to the lethal effects of endotoxin. Proc Natl Acad Sci U S A 76:5939–5943[CrossRef]
GolenbockD. T.,
HamptonR. Y.,
QureshiN.,
TakayamaK.,
RaetzC. R. H.
1991; Lipid A-like molecules that antagonize the effects of endotoxins on human monocytes. J Biol Chem 266:19490–19498
GreenL. C.,
WangerD. A.,
GlogowskyJ.,
SipperP. L.,
WishnokJ. S.,
TannenbaumS. R.
1982; Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem 126:131–138[CrossRef]
HommaJ. Y.,
MatsuuraM.,
KanegasakiS.11 other authors1985; Structural requirements of lipid A responsible for the functions: a study with chemically synthesized lipid A and its analogues. J Biochem 98:395–406
IidaT.,
HaishimaY.,
TanakaA.,
TanamotoK.
1996; Chemical structure of lipid A isolated from Comamonas testosteroni lipopolysaccharide. Eur J Biochem 237:468–475[CrossRef]
KanegasakiS.,
TanamotoK.,
YasudaT.10 other authors1986; Structure-activity relationship of lipid A: comparison of biological activities of natural and synthetic lipid A’s with different fatty acid compositions. J Biochem 99:1203–1210
KitchensR. L.,
UlevitchT. R. J.,
MunfordR. S.
1992; Lipopolysaccharide (LPS) partial structures inhibit responses to LPS in a human macrophage cell line without inhibiting LPS uptake by a CD14-mediated pathway. J Exp Med 176:485–494[CrossRef]
KotaniS.,
TakadaH.,
TakahashiI.7 other authors1986; Low endotoxic activities of synthetic Salmonella -type lipid A with an additional acyloxyacyl group on the 2-amino group of beta (1–6) glucosamine disaccharide 1–4′-bisphosphate. Infect Immun 52:872–884
KulshinV. A.,
ZaringerU.,
LindnerB.,
LagerK.,
DmitrievB. A.,
RietschelE. Th.
1991; Structural characterization of the lipid A component of Pseudomonas aeruginosa wild-type and rough mutant lipopolysaccharides. Eur J Biochem 198:697–704[CrossRef]
MayerH.,
WeckesserJ.
1984; ‘Unusual’ lipid A’s: structures, taxonomical relevance and potential value for endotoxin research. In Handbook of Endotoxins pp 221–241 Edited by
RietschelE. Th.
Amsterdam/New York/Oxford: Elsevier;
OgawaT.
1994; Immunobiological properties of chemically defined lipid A from lipopolysaccharide of Porphyromonas ( Bacteroides ) gingivalis . Eur J Biochem 219:737–742[CrossRef]
OmarA. S.,
FlammannH. T.,
BorowiakD.,
WeckesserJ.
1983; Lipopolysaccharide of two strains of the phototrophic bacterium Rhodopseudomonas capsulata. Arch Microbiol 134:212–216[CrossRef]
QureshiN.,
HonovichJ. P.,
HaraH.,
CotterR. J.,
TakayamaK.
1991; Diphosphoryl lipid A obtained from the nontoxic lipopolysaccharide of Rhodopseudomonas sphaeroides is an endotoxin antagonist in mice. Infect Immun 59:441–444
TanamotoK.
1999; Induction of lethal shock and tolerance by Porphyromonas gingivalis lipopolysaccharide in d-galactosamine-sensitized C3H/HeJ mice. Infect Immun 67:3399–3402
TanamotoK.,
AzumiS.
2000; Salmonella -type heptaacylated lipid A is inactive and acts as an antagonist of LPS action on human line cells. J Immunol 164:3149–3156[CrossRef]
TanamotoK.,
GalanosC.,
KusumotoS.,
ShibaT,
LüderitzO.1984; Mitogenic activities of chemically synthesized lipid A analogues and suppression of mitogenicity of lipid A. Infect Immun 44:427–433
TanamotoK.,
AzumiS.,
HaishimaY.,
KumadaH.,
UmemotoT.
1997a; The lipid A moiety of Porphyromonas gingivalis LPS specifically mediates the activation of C3H/HeJ mice. J Immunol 158:4430–4436
TroelstraA.,
Antal-SzalmasP.,
WeersinkA. J. L.,
VerhoefJ.,
Van KesselK. P. M.,
Van StrijpJ. A. G,
de Graff-MiltenburgL. A. M.1997; Saturable CD14-dependent binding of fluorescein-labeled lipopolysaccharide to human monocytes. Infect Immun 65:2272–2277
WestphalO.,
JannK.
1965; Bacterial lipopolysaccharides. Extraction with phenol-water and further applications of the procedure. Methods Carbohydr Chem 5:83–91