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Identification of FtsW as a transporter of lipid-linked cell wall precursors across the membrane.

Mohammadi T, van Dam V, Sijbrandi R, Vernet T, Zapun A, Bouhss A, Diepeveen-de Bruin M, Nguyen-Distèche M, de Kruijff B, Breukink E - EMBO J. (2011)

Bottom Line: The intracellular part of the pathway results in the production of the membrane-anchored cell wall precursor, Lipid II.The translocation (flipping) step of Lipid II was demonstrated to require a specific protein (flippase).This study provides the first biochemical evidence for the involvement of an essential protein in the transport of lipid-linked cell wall precursors across biogenic membranes.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical Biology and Organic Chemistry, Institute of Biomembranes, Bijvoet Center for Biomolecular Research, Faculty of Science, Utrecht University, Padualaan, Utrecht, The Netherlands.

ABSTRACT
Bacterial cell growth necessitates synthesis of peptidoglycan. Assembly of this major constituent of the bacterial cell wall is a multistep process starting in the cytoplasm and ending in the exterior cell surface. The intracellular part of the pathway results in the production of the membrane-anchored cell wall precursor, Lipid II. After synthesis this lipid intermediate is translocated across the cell membrane. The translocation (flipping) step of Lipid II was demonstrated to require a specific protein (flippase). Here, we show that the integral membrane protein FtsW, an essential protein of the bacterial division machinery, is a transporter of the lipid-linked peptidoglycan precursors across the cytoplasmic membrane. Using Escherichia coli membrane vesicles we found that transport of Lipid II requires the presence of FtsW, and purified FtsW induced the transbilayer movement of Lipid II in model membranes. This study provides the first biochemical evidence for the involvement of an essential protein in the transport of lipid-linked cell wall precursors across biogenic membranes.

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Related in: MedlinePlus

Schematic representation of the chemical structure of Lipid II (A) and NBD-labelled Lipid II (B). This peptidoglycan precursor consists of an undecaprenyl chain, phosphate (Pi), MurNAc (M) and GlcNAc (G). The pentapeptide moiety of MurNAc is symbolized by circles. NBD fluorophore is attached at the lysine of the pentapeptide moiety residue.
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f1: Schematic representation of the chemical structure of Lipid II (A) and NBD-labelled Lipid II (B). This peptidoglycan precursor consists of an undecaprenyl chain, phosphate (Pi), MurNAc (M) and GlcNAc (G). The pentapeptide moiety of MurNAc is symbolized by circles. NBD fluorophore is attached at the lysine of the pentapeptide moiety residue.

Mentions: The cell wall or peptidoglycan layer is unique and essential to bacteria. Most steps involved in its biosynthesis are therefore exploited as targets for well-known antibiotics or are explored for designing novel drugs. The central events in the building of peptidoglycan enclose the synthesis of two lipid intermediates on the cytoplasmic side of the membrane, Lipid I and Lipid II, and the subsequent transport of the latter across the bacterial membrane. In brief, the cytoplasmic step of peptidoglycan precursor synthesis culminates in the production of UDP-N-acetylmuramyl-pentapeptide (UDP-MurNAc-pentapeptide) from UDP-N-acetyl-glucosamine (UDP-GlcNAc). This compound is coupled to undecaprenyl phosphate to form Lipid I by MraY, an integral membrane protein. The subsequent addition of GlcNAc by MurG, an enzyme associated with the membrane, yields Lipid II (its structure is illustrated in Figure 1A). Thereafter, Lipid II is translocated to the exterior surface of the cell by an unknown mechanism and incorporated into the peptidoglycan through transglycosylation and transpeptidation reactions by penicillin-binding proteins (PBPs; Höltje, 1998; Cabeen and Jacobs-Wagner, 2007; Bouhss et al, 2008; den Blaauwen et al, 2008; Vollmer and Bertsche, 2008).


Identification of FtsW as a transporter of lipid-linked cell wall precursors across the membrane.

Mohammadi T, van Dam V, Sijbrandi R, Vernet T, Zapun A, Bouhss A, Diepeveen-de Bruin M, Nguyen-Distèche M, de Kruijff B, Breukink E - EMBO J. (2011)

Schematic representation of the chemical structure of Lipid II (A) and NBD-labelled Lipid II (B). This peptidoglycan precursor consists of an undecaprenyl chain, phosphate (Pi), MurNAc (M) and GlcNAc (G). The pentapeptide moiety of MurNAc is symbolized by circles. NBD fluorophore is attached at the lysine of the pentapeptide moiety residue.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3102273&req=5

f1: Schematic representation of the chemical structure of Lipid II (A) and NBD-labelled Lipid II (B). This peptidoglycan precursor consists of an undecaprenyl chain, phosphate (Pi), MurNAc (M) and GlcNAc (G). The pentapeptide moiety of MurNAc is symbolized by circles. NBD fluorophore is attached at the lysine of the pentapeptide moiety residue.
Mentions: The cell wall or peptidoglycan layer is unique and essential to bacteria. Most steps involved in its biosynthesis are therefore exploited as targets for well-known antibiotics or are explored for designing novel drugs. The central events in the building of peptidoglycan enclose the synthesis of two lipid intermediates on the cytoplasmic side of the membrane, Lipid I and Lipid II, and the subsequent transport of the latter across the bacterial membrane. In brief, the cytoplasmic step of peptidoglycan precursor synthesis culminates in the production of UDP-N-acetylmuramyl-pentapeptide (UDP-MurNAc-pentapeptide) from UDP-N-acetyl-glucosamine (UDP-GlcNAc). This compound is coupled to undecaprenyl phosphate to form Lipid I by MraY, an integral membrane protein. The subsequent addition of GlcNAc by MurG, an enzyme associated with the membrane, yields Lipid II (its structure is illustrated in Figure 1A). Thereafter, Lipid II is translocated to the exterior surface of the cell by an unknown mechanism and incorporated into the peptidoglycan through transglycosylation and transpeptidation reactions by penicillin-binding proteins (PBPs; Höltje, 1998; Cabeen and Jacobs-Wagner, 2007; Bouhss et al, 2008; den Blaauwen et al, 2008; Vollmer and Bertsche, 2008).

Bottom Line: The intracellular part of the pathway results in the production of the membrane-anchored cell wall precursor, Lipid II.The translocation (flipping) step of Lipid II was demonstrated to require a specific protein (flippase).This study provides the first biochemical evidence for the involvement of an essential protein in the transport of lipid-linked cell wall precursors across biogenic membranes.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical Biology and Organic Chemistry, Institute of Biomembranes, Bijvoet Center for Biomolecular Research, Faculty of Science, Utrecht University, Padualaan, Utrecht, The Netherlands.

ABSTRACT
Bacterial cell growth necessitates synthesis of peptidoglycan. Assembly of this major constituent of the bacterial cell wall is a multistep process starting in the cytoplasm and ending in the exterior cell surface. The intracellular part of the pathway results in the production of the membrane-anchored cell wall precursor, Lipid II. After synthesis this lipid intermediate is translocated across the cell membrane. The translocation (flipping) step of Lipid II was demonstrated to require a specific protein (flippase). Here, we show that the integral membrane protein FtsW, an essential protein of the bacterial division machinery, is a transporter of the lipid-linked peptidoglycan precursors across the cytoplasmic membrane. Using Escherichia coli membrane vesicles we found that transport of Lipid II requires the presence of FtsW, and purified FtsW induced the transbilayer movement of Lipid II in model membranes. This study provides the first biochemical evidence for the involvement of an essential protein in the transport of lipid-linked cell wall precursors across biogenic membranes.

Show MeSH
Related in: MedlinePlus