Open AccessChemical structure and immunoreactivity of the lipopolysaccharide of the deep rough mutant I‐69 Rd – /b + of Haemophilus influenzae
Author(s) -
MüllerLoennies Sven,
Brade Lore,
Brade Helmut
Publication year - 2002
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1033.2002.02763.x
Subject(s) - haemophilus influenzae , chemistry , lipid a , glucosamine , epitope , mutant , stereochemistry , acylation , nuclear magnetic resonance spectroscopy , lipopolysaccharide , oligosaccharide , biochemistry , antigen , biology , catalysis , genetics , gene , endocrinology , antibiotics
From the lipopolysaccharide of the deep rough mutant I‐69 Rd – /b + of Haemophilus influenzae two oligosaccharides were obtained after de‐ O ‐acylation and separation by high‐performance anion exchange chromatography. Their␣chemical structures were determined by one‐ and two‐dimensional 1 H‐, 13 C‐ and 31 P‐NMR spectroscopy as␣αKdo‐4 P ‐(2→6)‐βGlcN‐4 P ‐(1→6)‐αGlcN‐1 P and αKdo‐5 P ‐(2→6)‐βGlcN‐4 P ‐(1→6)‐αGlcN‐1 P . The specificity of mAbs S42‐21 and S42‐16 specific for Kdo‐4 P or Kdo‐5 P , respectively [Rozalski, A., Brade L., Kosma P., Moxon R., Kusumoto S., & Brade H. (1997). Mol. Microbiol . 23 , 569–577] was confirmed with neoglycoconjugates obtained by conjugation of the isolated oligosaccharides to BSA. In addition, a mAb S42‐10‐8 with unknown epitope specificity could be assigned using the neoglycoconjugates described herein. This mAb binds to an epitope composed of the bisphosphorylated glucosamine backbone of lipid A and Kdo‐4 P , whereby the latter determines the specificity strictly by the position of the phosphate group.