The lipooligosaccharide (LOS) of a Neisseria meningitidis acetate auxotroph was metabolically labeled with either [2-13C]-sodium acetate or [1,2-13C 2]-sodium acetate. In this study, we demonstrated that this label was efficiently incorporated into both the lipid A acyl moieties and the two N-acetylglucosamines present in the oligosaccharide branch of the LOS. The development of this efficient labeling protocol should prove useful in future structural studies analyzing the interactions between LOS and host proteins.
Harvey HA, Post DM, Apicella MAImmortalization of human urethral epithelial cells: a model for the study of the pathogenesis of and the inflammatory cytokine response to Neisseria gonorrhoeae infection. Infect Immun2002; 70: 5808—5815.
2.
Preston A., Mandrell RE, Gibson BW, Apicella MAThe lipooligosaccharides of pathogenic Gram-negative bacteria. Crit Rev Microbiol1996; 22: 139—180.
Raetz CRBacterial endotoxins: extraordinary lipids that activate eucaryotic signal transduction. J Bacteriol1993; 175: 5745—5753.
5.
Sunshine MG, Gibson BW, Engstrom JJ, Nichols WA, Jones BD, Apicella MAMutation of the htrB gene in a virulent Salmonella typhimurium strain by intergeneric transduction: strain construction and phenotypic characterization. J Bacteriol1997; 179: 5521—5533.
6.
Lee NG, Sunshine MG, Engstrom JJ, Gibson BW, Apicella MAMutation of the htrB locus of Haemophilus influenzae nontypable strain 2019 is associated with modifications of lipid A and phosphorylation of the lipo—oligosaccharide . J Biol Chem1995; 270: 27151—27159.
7.
Kulshin VA, Zahringer U., Lindner B. et al. Structural characterization of the lipid A component of pathogenic Neisseria meningitidis. J Bacteriol1992; 174: 1793—1800.
8.
Rahman MM, Stephens DS, Kahler CM, Glushka J., Carlson RWThe lipooligosaccharide (LOS) of Neisseria meningitidis serogroup B strain NMB contains L2, L3, and novel oligosaccharides, and lacks the lipid-A 4′—phosphate substituent. Carbohydr Res1998; 307: 311—324.
9.
Rietschel ET , Kirikae T., Schade FU et al. Bacterial endotoxin: molecular relationships of structure to activity and function. FASEB J1994; 8: 217—225.
10.
Teghanemt A. , Zhang D., Levis DN, Weiss JP, Gioannini TLMolecular basis of reduced potency of under-acylated endotoxins. J Immunol2005; 175: 4669—4676.
11.
Unkmeir A., Kammerer U., Stade A. et al. Lipooligosaccharide and polysaccharide capsule: virulence factors of Neisseria meningitidis that determine meningococcal interaction with human dendritic cells. Infect Immun2002; 70: 2454—2462.
12.
Bjerre A., Brusletto B., Rosenqvist E. et al. Cellular activating properties and morphology of membrane-bound and purified meningococcal lipopolysaccharide. J Endotoxin Res2000; 6: 437—445.
13.
Beutler B., Rietschel ETInnate immune sensing and its roots: the story of endotoxin. Nat Rev Immunol2003; 3:169—176.
14.
Gioannini TL , Teghanemt A., Zarember KA, Weiss JPRegulation of interactions of endotoxin with host cells. J Endotoxin Res2003; 9: 401—408.
15.
Gioannini TL , Teghanemt A., Zhang D. et al. Isolation of an endotoxin—MD-2 complex that produces Toll-like receptor 4-dependent cell activation at picomolar concentrations. Proc Natl Acad Sci USA2004 ; 101: 4186-419 1.
16.
Miyake K.Innate recognition of lipopolysaccharide by CD14 and Toll-like receptor 4-MD-2: unique roles for MD-2. Int Immunopharmacol2003; 3: 119—128.
17.
Ulevitch RJ , Tobias PSRecognition of Gram-negative bacteria and endotoxin by the innate immune system. Curr Opin Immunol1999; 11: 19—22.
18.
Gioannini TL , Teghanemt A., Zhang D., Levis EN, Weiss JPMonomeric endotoxin:protein complexes are essential for TLR4-dependent cell activation. J Endotoxin Res2005; 11: 117—123.
19.
Brandtzaeg P., Bjerre A., Ovstebo R., Brusletto B., Joo GB, Kierulf P.Neisseria meningitidis lipopolysaccharides in human pathology. J Endotoxin Res2001; 7: 401—408.
20.
Giardina PC , Gioannini T., Buscher BA et al. Construction of acetate auxotrophs of Neisseria meningitidis to study host-meningococcal endotoxin interactions . J Biol Chem2001; 276: 5883—5891.
21.
Post DM, Phillips NJ, Shao JQ, Entz DD, Gibson BW, Apicella MAIntracellular survival of Neisseria gonorrhoeae in male urethral epithelial cells: importance of a hexaacyl lipid A. Infect Immun2002; 70: 909—920.
22.
Kahler CM, Datta A., Tzeng YL, Carlson RW, Stephens DSInner core assembly and structure of the lipooligosaccharide of Neisseria meningitidis: capacity of strain NMB to express all known immunotype epitopes . Glycobiology2005; 15: 409—419.
23.
Mengin-Lecreulx D., van Heijenoort J.Copurification of glucosamine-1-phosphate acetyltransferase and N-acetylglucosamine-1-phosphate uridyltransferase activities of Escherichia coli: characterization of the glmU gene product as a bifunctional enzyme catalyzing two subsequent steps in the pathway for UDP-N-acetylglucosamine synthesis. J Bacteriol1994; 176: 5788—5795.
24.
Mengin-Lecreulx D., van Heijenoort J.Identification of the glmU gene encoding N-acetylglucosamine-1-phosphate uridyltransferase in Escherichia coli. J Bacteriol1993; 175 : 6150—6157.