NMR Based Structural Analysis of the Complete Rough-type LPS Isolated From Capnocytophaga canimorsus

Zähringer, Ulrich and Ittig, Simon and Lindner, Buko and Moll, Hermann and Schombel, Ursula and Gisch, Nicolas and Cornelis, Guy R.. (2014) NMR Based Structural Analysis of the Complete Rough-type LPS Isolated From Capnocytophaga canimorsus. Journal of biological chemistry, Vol. 289, H. 34. pp. 23963-23976.

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Official URL: http://edoc.unibas.ch/dok/A6288856

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We here describe the NMR analysis of an intact lipopolysaccharide (LPS, endotoxin) in water with 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) as detergent. When HPLC-purified rough-type LPS of Capnocytophaga canimorsus was prepared, 13C,15N-labelling could be avoided. The intact LPS was analyzed by homo- (1H) and heteronuclear (1H,13C, and 1H,31P) correlated 1D and 2D NMR techniques as well as by mass spectrometry. It consists of a penta-acylated lipid A with an α-linked phospho-ethanolamine (P-Etn) attached to C-1 of GlcN (I) in the hybrid backbone, lacking the 4′-phosphate. The hydrophilic core-oligosaccharide was found to be a complex hexasaccharide with two Man and one of each Kdo, Gal, GalN, and L-Rha residues. Position 4 of Kdo is substituted by P-Etn, also present in position 6 of the branched ManI residue. This rough-type LPS is exceptional in that all three negative phosphate residues are 'masked' by positively charged ethanolamine substituents leading to an overall zero net charge, which has so far not been observed for any other LPS. In biological assays the corresponding isolated lipid A was found to be endotoxically almost inactive. By contrast, the intact rough type LPS described here expressed a 20,000 fold increased endotoxicity, indicating that the core-oligosaccharide significantly contributes to the endotoxic potency of the whole rough-type C. canimorsus LPS molecule. Based on these findings, the strict view that lipid A alone represents the toxic center of LPS needs to be reassessed.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Former Organization Units Biozentrum > Molecular Microbiology (Cornelis)
UniBasel Contributors:Ittig, Simon Josef
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:American Society of Biological Chemists
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:10 Oct 2014 09:19
Deposited On:10 Oct 2014 09:19

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