Mode of elongation of the glycerol phosphate polymer of membrane lipoteichoic acid of Streptococcus faecium ATCC 9790

Cabacungan, E; Pieringer, Ronald A.

doi:10.1128/jb.147.1.75-79.1981

Cited by 31 publications

(6 citation statements)

References 18 publications

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“…Three observations suggested that inorganic phosphate and other anions stimulate one or more reactions in the assembly of lipophilic and hydrophilic LTA in toluene-treated cells of L. casei. First, inorganic phosphate greatly increased the incorporation of [ tion supports the conclusion of Cabacungan and Pieringer (5), who reported that GroP units were added distally in whole cells and isolated membranes of Streptococcus faecium. Thus, the mode of distal addition of GroP to LTA in toluene-treated cells of L. casei appears to be similar to that in S. faecium cells.…”

Section: Resultssupporting

confidence: 74%

Biosynthesis of D-alanyl-lipoteichoic acid: role of diglyceride kinase in the synthesis of phosphatidylglycerol for chain elongation

Taron¹,

rd²,

Neuhaus³

1983

J Bacteriol

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Lipophilic and hydrophilic D-alanyl-lipoteichoic acids are elongated in Lactobacillus casei by the transfer of sn-glycerol 1-phosphate units from phosphatidylglycerol to the poly(glycerophosphate) moiety of the polymer. These sn-glycerol 1phosphate units are added to the end of the poly(glycerophosphate) which is distal to the glycolipid anchor; 1,2-diglyceride results from this addition. The presence of a diglyceride kinase was suggested by the ATP-dependent phosphorylation of 1,2-diglyceride to phosphatidic acid. Inorganic phosphate was used to initiate the synthesis of lipophilic lipoteichoic acid (LTA) and the elongation of both lipophilic and hydrophilic LTA. Three observations suggest that phosphate and other anions play a role in the in vitro synthesis of LTA and its precursors. First, the conversion of 1,2-diglyceride to phosphatidic acid by diglyceride kinase was stimulated. Second, the synthesis of phosphatidylglycerol was increased. Third, the elongation of lipophilic and hydrophilic LTA was enhanced. These observations indicated that one effect of phosphate might be to enhance the utilization of 1,2-diglyceride for the synthesis of phosphatidic acid. This phospholipid is a precursor of phosphatidylglycerol, the donor of sn-glycerol 1-phosphate for elongation of LTA. Phosphatidylglycerol (PG) has been proposed to be the donor of sn-glycerol 1-phosphate (GroP) units of the poly(glycerophosphate) moiety of lipoteichoic acid (LTA) (8,9,13,14) according to the following reaction: tion would result in a cycling of the diglyceride moiety of this phospholipid.The goals of these experiments with Lactobacillus casei were to characterize further the effect of phosphate on the assembly of the LTA, PG + LTA-poly(glycerophosphate)" -* LTA-poly(glycerophosphate),+1 + 1,2-diglyceride Inorganic phosphate stimulates both the synthesis of PG and the elongation of D-alanyl-lipophilic LTA in vitro (4). These observations supported the proposed role for PG in the elongation of LTA.During the elongation of LTA, one of the reaction products is 1,2-diglyceride. Significant amounts of this diglyceride might be expected to accumulate during chain elongation. Since large amounts of diglyceride are not commonly found in bacteria (25), it is proposed that the diglyceride is either degraded or reutilized for phospholipid synthesis. A diglyceride kinase similar to that found in Escherichia coli (24) could phosphorylate the diglyceride to phosphatidic acid, a known precursor of PG (25). This phosphorylat Present address: College of Dentistry, University of Illinois at the Medical Center, Chicago, IL 60612. to determine the site of the addition of GroP units to the growing polymer, and to suggest a fate for the 1,2-diglyceride. Toluene-treated cells were used to demonstrate the synthesis and elongation of D-alanyl-LTA as well as the synthesis of various phospholipids. These cells, which are permeable to GroP, ATP, and Dalanine, synthesized LTA and phospholipids in significant amounts. For detecting diglyceride kinase, membranes were u...

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Section: Resultssupporting

confidence: 74%

Biosynthesis of D-alanyl-lipoteichoic acid: role of diglyceride kinase in the synthesis of phosphatidylglycerol for chain elongation

Taron¹,

rd²,

Neuhaus³

1983

J Bacteriol

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“…The LtaS membrane domain likely contains binding sites for both the lipid anchor and for PG, which serves as the phosphoglycerol donor. Because LTA elongates from the growing end (27,28), and (as our data show) the released polymers cannot productively rebind, there must also be a region in the extracellular domain that holds the end of the growing chain. To identify potential lipid-binding sites in the membrane domain, we analyzed sequence conservation (48) and identified two clusters of well-conserved residues, one directly adjacent to the active site of the extracellular domain in our model and the other more distal from the active site and extracellular domain (SI Appendix, Fig.…”

Section: A Model For Polymer Length Control By Starter Unit Concentrationmentioning

confidence: 57%

“…The polymer elongates in a process that involves the repeated transfer of phosphoglycerol units from phosphatidylglycerol (PG) to a catalytic threonine in LtaS (T300) and then to the tip of the growing polymer (Fig. 1B) (27)(28)(29). Repeat units may be modified by D-alanyl esters or, less commonly, GlcNAc moieties (24,30).…”

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confidence: 99%

Lipoteichoic acid polymer length is determined by competition between free starter units

Hesser

Schaefer

Lee

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Carbohydrate polymers exhibit incredible chemical and structural diversity, yet are produced by polymerases without a template to guide length and composition. As the length of carbohydrate polymers is critical for their biological functions, understanding the mechanisms that determine polymer length is an important area of investigation. Most Gram-positive bacteria produce anionic glycopolymers called lipoteichoic acids (LTA) that are synthesized by lipoteichoic acid synthase (LtaS) on a diglucosyl-diacylglycerol (Glc2DAG) starter unit embedded in the extracellular leaflet of the cell membrane. LtaS can use phosphatidylglycerol (PG) as an alternative starter unit, but PG-anchored LTA polymers are significantly longer, and cells that make these abnormally long polymers exhibit major defects in cell growth and division. To determine how LTA polymer length is controlled, we reconstitutedStaphylococcus aureusLtaS in vitro. We show that polymer length is an intrinsic property of LtaS that is directly regulated by the identity and concentration of lipid starter units. Polymerization is processive, and the overall reaction rate is substantially faster for the preferred Glc2DAG starter unit, yet the use of Glc2DAG leads to shorter polymers. We propose a simple mechanism to explain this surprising result: free starter units terminate polymerization by displacing the lipid anchor of the growing polymer from its binding site on the enzyme. Because LtaS is conserved across most Gram-positive bacteria and is important for survival, this reconstituted system should be useful for characterizing inhibitors of this key cell envelope enzyme.

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“…In Streptococcus faecium the elongation of LTA is accomplished by the distal addition of sn-glycerol 1phosphate from phosphatidylglycerol (Cabacugan & Pieringer, 1981). This mode of' addition was also established in L. casei from the gradient of label generated in the poly(g1ycerophosphate) chain when toluene-treated cells were incubated with labelled glycerol 3-phosphate in the presence of 100 mM-inorganic phosphate (Taron et al, 1983).…”

mentioning

confidence: 88%

Inter-chain transacylation of d-alanine ester residues of lipoteichoic acid: a unique mechanism of membrane communication

Neuhaus

1985

Biochemical Society Transactions

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Lipoteichoic acid is an important example of an amphiphile that appears to play a vital role in bacterial growth. A number of functions have been ascribed to this polymer. These include the regulation of autolytic enzymes, the binding of Mgz+ for enzyme function, and the assembly of wall polymers. One of the key components which has been proposed to modulate these functions is the substituent D-alanine. James Baddiley and his co-workers discovered that this amino acid is an important component of most teichoic acids (Armstrong et al., 1959). Thus, esterification of teichoic acids by D-alanine may be an important step in controlling the function of this polymer in wall assembly. In Lactobacillus casei the incorporation of D-alanine into lipoteichoic acid is accomplished in a two-step reaction sequence. The first step of this sequence was discovered in 1960 when Baddiley & Neuhaus showed that a variety of Gram-positive organisms have the ability to activate Dalanine. The second step of this sequence was not detected until Reusch & Neuhaus (1971) recognized that the incorporation of D-alanine into teichoic acids in vitro required the endogenous membrane teichoic acid to act as acceptor. All attempts to demonstrate the incorporation of D-alanine into isolated wall teichoic acid proved to be elusive and it was reasoned that the teichoic acid has to be intimately associated with the membrane in a particular conformation or environment (Neuhaus et aZ., 1974). On the basis of this hypothesis a system for the incorporation of D-alanine into membranes was found that required both a supernatant fraction and ATP, and was stimulated by metal ions. Linzer & Neuhaus (1973) showed that the supernatant fraction could be replaced by the D-alanine-activating enzyme and a protein factor designated D-alanine: membrane acceptor ligase. L. casei was chosen for these experiments because, firstly, it has a high concentration of the Abbreviation used: LTA, lipoteichoic acid. *Dedicated to Sir James Baddiley.

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Mode of elongation of the glycerol phosphate polymer of membrane lipoteichoic acid of Streptococcus faecium ATCC 9790

Cited by 31 publications

References 18 publications

Biosynthesis of D-alanyl-lipoteichoic acid: role of diglyceride kinase in the synthesis of phosphatidylglycerol for chain elongation

Biosynthesis of D-alanyl-lipoteichoic acid: role of diglyceride kinase in the synthesis of phosphatidylglycerol for chain elongation

Lipoteichoic acid polymer length is determined by competition between free starter units

Inter-chain transacylation of d-alanine ester residues of lipoteichoic acid: a unique mechanism of membrane communication

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