Separation of the poly(glycerophosphate) lipoteichoic acids of Enterococcus faecalis Kiel 27738, Enterococcus hirae ATCC 9790 and Leuconostoc mesenteroides DSM 20343 into molecular species by affinity chromatography on concanavalin A

Abstract

This study shows for the first time microheterogeneity of 1,3-1inked poly(g1ycerophosphate) lipoteichoic acids. The lipoteichoic acids investigated were those of Enterococcus faecalis Kiel27738 (I), Enterococcus hirae (Streptococcusfaecium) ATCC 9790 (11), and Leuconostoc mesenteroides DMS 20343 (111). Lipoteichoic acids I1 and 111 are partially substituted by mono-, di-, tri-, and tetra-a-D-glUCOpyranOSyl residues with (1 + 2) interglycosidic linkages. Lipoteichoic acid I is substituted with a-kojibiosyl residues only. Lipoteichoic acids I and I11 additionally carry D-alanine ester.Lipoteichoic acids were separated on columns of concanavalin-A -Sepharose according to their increasing number of glycosyl substituents per chain. It was evident that all molecular species are usually glycosylated and that alanine ester and glycosyl residues occur on the same chains. The chain lengths of lipoteichoic acid I and I1 vary between 9 -40 glycerophosphate residues, whereas those of lipoteichoic acid I11 appear to be uniform (33 f 2 residues). Molecular species differ in the extent of glycosylation but their content of alanyl residues is fairly constant. All lipoteichoic acids contain a small fraction ( 5 -15%) different in composition from the bulk and most likely reflecting an early stage of biosynthesis.Two procedures for chain length determination of poly(g1ycerophosphate) lipoteichoic acids are described.Lipoteichoic acids are components of the cytoplasmic membrane of numerous Gram-positive bacteria [l]. Most widespread is the classical type which contains a single unbranched 1,3-1inked poly(g1ycerophosphate) chain consistently attached by a phosphodiester bond to C-6 of the nonreducing hexosyl terminus of a di-or trihexosyldiacylglycerol [l, 21. Some of the glycerol residues of the chain are substituted with glycosyl residues or D-alanine ester or with both. These chain substituents have the capacity to greatly modify the biological activities of lipoteichoic acids. D-Alanine ester and glycosyl substituents block the carrier activity of lipoteichoic acid in the reaction of wall teichoic acid polymerases [3, 41. DAlanine ester modifies the inhibitory effect of autolysins [5] and both substituents drastically reduce the ability to activate the mammalian complement system [6].Although poly(g1ycerophosphate) lipoteichoic acids were discovered 20 years ago [7, 81, a number of structural details has remained unexplored. Lipoteichoic acids of various bacteria contain alanine ester and glycosyl substituents but whether the two substituents occur on separate chains or together on the same chain has not previously been clarified. Since substitution is commonly incomplete, another question is whether these lipoteichoic acids are composed of fully sub- Answers to all three questions are presented in this report in which we describe affinity chromatography of lipoteichoic acids on concanavalin A, a lectin of the jack bean [14]. To be recognized by this lectin, lipoteichoic acids were selected that carry a-D-glUCOpyranOSyl and (...

Role of Lipoteichoic Acid in the Phagocyte Response to Group BStreptococcus

“…Furthermore, the structure of the glycolipid Glc-␣-1,2Glc-␣-1-3-diacylglycerol was different from those revealed in S. aureus and S. pneumoniae but similar to those in Enterococcus, Leuconostoc, and Lactococcus spp. (36). In addition, in GBS LTA the polyglycerophosphate backbone was substituted with D-alanine only whereas in S. aureus and many other species D-alanine substituents are alternating with N-acetyl-glucosamine.…”

A Continuum of Anionic Charge: Structures and Functions ofd-Alanyl-Teichoic Acids in Gram-Positive Bacteria

“…Distal addition can occur only while the growing poly(Gro-P) moiety is in contact with the cytoplasmic membrane. The polydispersity of the LTA chain length suggests that chain growth may terminate at any point in elongation (300). Comparative studies of LTA glycosylation in B. subtilis further illustrate aspects of structural diversity.…”

The Sweet Tooth of Bacteria: Common Themes in Bacterial Glycoconjugates

“…The glycosylation of LTA can be used as a taxonomic marker for S. sanguis, as its (1-6)-bound mono-, di-, tri-, and tetra-␣-Dglucosyl structures are unique (258)(259)(260). The LTA of Enterococcus species is similarly decorated with sugar oligomers, but these are linked to the backbone via a (1,2)-bound structure (261). The glycosylation of TAs has also been reported for some lactic acid bacteria such as L. plantarum (Glc on WTA) (236,262) and Lactococcus lactis subsp.…”