Document Detail

Epitope mapping of the O-chain polysaccharide of Legionella pneumophila serogroup 1 lipopolysaccharide by saturation-transfer-difference NMR spectroscopy.
MedLine Citation:
PMID:  11856316     Owner:  NLM     Status:  MEDLINE    
Two modifications of 5-acetimidoylamino-7-acetamido-3,5,7,9-tetradeoxy-D-glycero-D-galacto-non-2-ulosonic acid (5-N-acetimidoyl-7-N-acetyllegionaminic acid) in the O-chain polysaccharide (OPS) of the Legionella pneumophila serogroup 1 lipopolysaccharide (LPS) concern N-methylation of the 5-N-acetimidoyl group in legionaminic acid. Both N-methylated substituents, the (N,N-dimethylacetimidoyl) amino and acetimidoyl(N-methyl)amino group, could be allocated to one single legionaminic acid residue in the long- and middle-chain OPS, respectively. Using mutants devoid of N-methylated legionaminic acid derivatives, it could be shown that N-methylation of legionaminic acid correlated with the expression of the mAb 2625 epitope. In the present study we investigated the binding of the LPS-specific monoclonal antibody mAb 2625 to isolated OPS with surface-plasmon-resonance biomolecular interaction analysis and saturation-transfer-difference (STD) NMR spectroscopy in order to map the mAb 2625 epitope on a molecular level. It could be demonstrated that the binding affinity of the N-methylated legionaminic acid derivatives was independent from the size of the isolated OPS molecular species. In addition, STD NMR spectroscopic studies with polysaccharide ligands with an average molecular mass of up to 14 kDa revealed that binding was mainly mediated via the N-methylated acetimidoylamino group and via the closely located 7-N-acetyl group of the respective legionaminic acid residue, thus indicating these derivatives to represent the major epitope of mAb 2625.
Oliver Kooistra; Lars Herfurth; Edeltraud Lüneberg; Matthias Frosch; Thomas Peters; Ulrich Zähringer
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  European journal of biochemistry / FEBS     Volume:  269     ISSN:  0014-2956     ISO Abbreviation:  Eur. J. Biochem.     Publication Date:  2002 Jan 
Date Detail:
Created Date:  2002-02-21     Completed Date:  2002-03-19     Revised Date:  2007-07-23    
Medline Journal Info:
Nlm Unique ID:  0107600     Medline TA:  Eur J Biochem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  573-82     Citation Subset:  IM    
Research Center Borstel, Center for Medicine and Biosciences, Germany.
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MeSH Terms
Antibodies, Monoclonal / chemistry
Carbohydrate Conformation
Carbohydrate Sequence
Epitope Mapping*
Hydrogen-Ion Concentration
Legionella pneumophila / chemistry*
Molecular Sequence Data
Nuclear Magnetic Resonance, Biomolecular
O Antigens / chemistry*
Surface Plasmon Resonance
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Ligands; 0/O Antigens

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