The inner core region of Yersinia enterocolitica Ye75R (0:3) lipopolysaccharide

Abstract

The inner-core region of the lipopolysaccharide of an UDPGalNAc-4-epimerase-deficient mutant of Yersinia enterocolitica 0 : 3, designated as Ye75R, was investigated using methylation analysis, 1 D-"C-NMR and 2D-"C-NMR and 'H-NMR, as well as 31P-NMR, fast-atom-bombardment mass spectrometry (FAB MS) and FAB M S N S in positive and negative modes. The isolated core heptasaccharide (0s) was composed of 2 units D-glucose, 3 units LD-heptose and 1 unit each of DD-heptose and 3-deoxy-~-manno-octulosonic acid. Methylation analysis indicated that 0s was highly branched with terminal location of the two glucoses and the DD-heptose unit, which was partially (to about 40%) phosphorylated at C7. These combined studies allowed us to formulate the structure of the inner core region as shown in Scheme 1.The substitution of the 7-position of the terminally linked DD-heptose unit by phosphate could be recognized by MS characterization of permethylated ~~-heptose-7-phosphate (alditol acetate) and the extent of the substitution by the ratio of the two well separated 'H signals of DD-heptose in 500-MHz 'H-NMR. Negative FAB MS of 0s also indicated the presence of smaller amounts of two hexasaccharides, differing from 0s in lacking either one terminal unit of D-glucose or of the tenninal DD-heptose, and additionally of a pentasaccharide lacking two heptosyl units, namely the terminal DD-heptose and and the subterminal LD-heptose. The presence of the smaller oligosaccharides in the 0s fraction was also recognized by the methylation analysis.Structural studies on the R-core region of a number of enterobacterial lipopolysaccharides (LPS) and of LPS of some phylogenetically rather remote bacterial species have been carried out in the past [l, 21. The core region, together with the lipid A component, are the two structural regions which are present in almost all Gram-negative bacteria and, as such, are of interest especially with regard to their immunoreactive properties [l]

The inner-core lipopolysaccharide biosynthetic waaE gene: function and genetic distribution among some Enterobacteriaceae b bThe GenBank accession number for the waaE gene sequences of P. mirabilis CECT170, Y. enterocolitica R102 and Ent. aerogenes CECT684 reported in this paper are AY075039, AY075041 and AY075040, respectively.

“…marcescens waaE mutant strains, thus proving that all the waaE homologues perform the same function in inner-core LPS biosynthesis. These results agree with core LPS structural studies in P. mirabilis and Y. enterocolitica where a substitution of ,-HeppI at the O-4 position by a Glcp [β--Glcp-(1 4)-α-,-HeppI] has been found (Radziejewska- Lebrecht et al, 1994 ;Vinogradov et al, 2000). From the Enterobacteriaceae waa gene cluster distal end sequence data a set of primers was designed to amplify genomic DNA containing the 3h end of the waaA gene up to the 3h end of the fpg gene.…”

“…In other species, like Klebsiella pneumoniae, a substitution of ,-HeppI at the O-4 position by a β--glucopyranose residue (β--Glcp) is found (Vinogradov & Perry, 2001). Furthermore, in K. pneumoniae, Proteus mirabilis and Yersinia enterocolitica a substitution of ,-HeppII at the O-3 position by an α--galacturonic acid residue (α--GalpA) residue is found (Radziejewska-Lebrecht et al, 1994 ;Vinogradov et al, 2000 ;Vinogradov & Perry, 2001), while a substitution of ,-HeppII at the O-3 position by a Glcp residue is found in E. coli (Heinrichs et al, 1998).…”

“…It consists of three domains : lipid A, core L. Izquierdo and others structure contains two residues of 3-deoxy--mannooctulopyranosonic acid (Kdop) and three residues of -glycero--manno-heptopyranose (,-HeppI, ,-HeppII and ,-HeppIII) (Radziejewska-Lebrecht et al, 1994 ;Heinrichs et al, 1998 ;Vinogradov et al, 2000 ;Vinogradov & Perry, 2001). In addition, some Enterobacteriaceae, like E. coli, contain a phosphoryl group modification at the ,-HeppI residue (Heinrichs et al, 1998).…”

Identification of a Novel Heptoglycan of α1→2-Linkedd-glycero-d-manno-Heptopyranose

“…pneumoniae R20 contains a heptaacyl component in varying amounts, depending on the growth conditions (data not shown). The structure resembles that of lipid A from K. pneumoniae O3 (48); however, as proven by 31 P NMR spectroscopy of de-Oacylated LPS and by the bisphosphorylated products obtained after alkaline treatment of de-O-acylated LPS (4), it lacks the 4-amino-4-deoxy-L-arabinose that substitutes the phosphate residue at O-4Ј in lipid A from K. pneumoniae O3. The structure of lipid A from K. pneumoniae ssp.…”

Enterobacterial common antigen and O-specific polysaccharide coexist in the lipopolysaccharide of Yersinia enterocolitica serotype O : 3

“…The structure of Y. enterocolitica O : 3 LPS has been determined (Hoffman et al, 1980;Holst, 2007;Pinta et al, 2009;Radziejewska-Lebrecht et al, 1994Shashkov et al, 1995) and proved to be somewhat unorthodox (Fig. 1).…”