title: Chemical structure and immunobiological activity of lipid A from Serratia marcescens LPS.
abstract: The chemical structure and immunobiological activities of Serratia marcescens lipid A, an active centre of LPS, were investigated. LPS preparations of S. marcescens were extracted using a hot phenol/water method, after which purified lipid A specimens were prepared by weak acid hydrolysis, followed by normal phase and gel filtration chromatographic separation. The lipid A structure was determined by MS to be a diglucosamine backbone with diphosphates and five C(14) normal chain acyl groups, including two acyloxyacyl groups at the 2 and 3 positions of the non-reducing side. S. marcescens lipid A and Escherichia coli-type synthetic lipid A (compound 506) exhibited definite reactivity in Limulus amoebocyte lysate assays. The lethal toxicity of S. marcescens lipid A was nearly comparable to that of compound 506, and both induced nuclear factor-kappaB activation in murine cells via Toll-like receptor (TLR)4/MD-2 but not TLR2, as well as various inflammatory cytokines in peritoneal macrophages of C3H/HeN mice but not C3H/HeJ mice. Furthermore, S. marcescens lipid A induced nearly the same amounts of tumour necrosis factor alpha, interleukin-6, and nitric oxide production by the murine alveolar macrophage cell line MH-S as compared with compound 506. These results indicate that S. marcescens possesses a penta-acylated lipid A, which is nearly identical to E. coli lipid A in regard to biological activities, while it also may be a crucial virulence factor of the bacterium.
authors.affiliation: Department of Oral Microbiology, Asahi University School of Dentistry, 1851-1 Hozumi, Mizuho, Gifu 501-0296, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Makimura
authors.affiliation: Department of Oral Microbiology, Asahi University School of Dentistry, 1851-1 Hozumi, Mizuho, Gifu 501-0296, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Asai
authors.affiliation: Department of Oral Microbiology, Asahi University School of Dentistry, 1851-1 Hozumi, Mizuho, Gifu 501-0296, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Sugiyama
authors.affiliation: Department of Oral Microbiology, Asahi University School of Dentistry, 1851-1 Hozumi, Mizuho, Gifu 501-0296, Japan.
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Ogawa
mesh_terms: D000818:Animals; D005591:Chemical Fractionation / Q000379:methods; D002850:Chromatography, Gel; D002853:Chromatography, Liquid; D016207:Cytokines / Q000096:biosynthesis; D008033:Limulus Test; D008050:Lipid A / Q000737:chemistry* / Q000276:immunology* / Q000633:toxicity; D008070:Lipopolysaccharides / Q000737:chemistry / Q000302:isolation & purification; D016676:Macrophages, Alveolar / Q000276:immunology; D017737:Macrophages, Peritoneal / Q000276:immunology; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D015394:Molecular Structure; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D011041:Poisoning / Q000401:mortality; D012706:Serratia marcescens / Q000737:chemistry* / Q000276:immunology*; D051195:Toll-Like Receptor 2 / Q000276:immunology; D051197:Toll-Like Receptor 4 / Q000276:immunology
chemical_list: D016207:Cytokines; D008050:Lipid A; D008070:Lipopolysaccharides; D016328:NF-kappa B; C493486:Tlr2 protein, mouse; C493487:Tlr4 protein, mouse; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D009569:Nitric Oxide