scholarly journals Lipid Flippases for Bacterial Peptidoglycan Biosynthesis

2015 ◽  
Vol 8s1 ◽  
pp. LPI.S31783 ◽  
Author(s):  
Natividad Ruiz

The biosynthesis of cellular polysaccharides and glycoconjugates often involves lipid-linked intermediates that need to be translocated across membranes. Essential pathways such as N-glycosylation in eukaryotes and biogenesis of the peptidoglycan (PG) cell wall in bacteria share a common strategy where nucleotide-sugars are used to build a membrane-bound oligosaccharide precursor that is linked to a phosphorylated isoprenoid lipid. Once made, these lipid-linked intermediates must be translocated across a membrane so that they can serve as substrates in a different cellular compartment. How translocation occurs is poorly understood, although it clearly requires a transporter or flippase. Identification of these transporters is notoriously difficult, and, in particular, the identity of the flippase of lipid II, an intermediate required for PG biogenesis, has been the subject of much debate. Here, I will review the body of work that has recently fueled this controversy, centered on proposed flippase candidates FtsW, MurJ, and AmJ.

2013 ◽  
Vol 57 (9) ◽  
pp. 4267-4275 ◽  
Author(s):  
Yong Heon Lee ◽  
John D. Helmann

ABSTRACTUndecaprenyl pyrophosphate synthase (UppS) catalyzes the formation of the C55lipid carrier (UPP) that is essential for bacterial peptidoglycan biosynthesis. We selected here a vancomycin (VAN)-resistant derivative ofBacillus subtilisW168 that contains a single-point mutation in the ribosome-binding site of theuppSgene designateduppS1. Genetic reconstruction experiments demonstrate that theuppS1allele is sufficient to confer low-level VAN resistance and causes reduced UppS translation. The decreased level of UppS rendersB. subtilisslightly more susceptible to many late-acting cell wall antibiotics, including β-lactams, but significantly more resistant to fosfomycin andd-cycloserine, antibiotics that interfere with the very early steps of cell wall synthesis. We further show that theuppS1allele leads to slightly elevated expression of the σMregulon, possibly helping to compensate for the stress caused by a decrease in UPP levels. Notably, theuppS1mutation increases resistance to VAN, fosfomycin, andd-cycloserine in wild-type cells, but this effect is greatly reduced or eliminated in asigMmutant background. Our findings suggest that, although UppS is an attractive antibacterial target, incomplete inhibition of UppS function may lead to increased resistance to some cell wall-active antibiotics.


2019 ◽  
Author(s):  
Alvin C. Y. Kuk ◽  
Aili Hao ◽  
Ziqiang Guan ◽  
Seok-Yong Lee

AbstractThe biosynthesis of many polysaccharides, including bacterial peptidoglycan and eukaryotic N-linked glycans, requires transport of lipid-linked oligosaccharide (LLO) precursors across the membrane by specialized flippases. MurJ is the flippase for the lipid-linked peptidoglycan precursor Lipid II, a key player in bacterial cell wall synthesis, and a target of recently discovered antibacterials. However, the flipping mechanism of LLOs including Lipid II remains poorly understood due to a dearth of structural information. Here we report crystal structures of MurJ captured in inward-closed, inward-open, inward-occluded and outward-facing conformations. Together with cysteine accessibility, mass spectrometry, and complementation studies, we elucidate the conformational transitions in MurJ that mediate lipid flipping, identified the key ion for function, and provide a framework for the development of inhibitors.


2021 ◽  
Author(s):  
Atsushi Taguchi ◽  
Suzanne Walker

ABSTRACTThe peptidoglycan cell wall is a macromolecular structure that encases bacteria and is essential for their survival. Proper assembly of the cell wall requires peptidoglycan synthases as well as membrane-bound cleavage enzymes that control where new peptidoglycan is made and inserted. We are only beginning to understand the roles of peptidoglycan cleavage enzymes in cell wall assembly. Previous studies have shown that two membrane-bound proteins in Streptococcus pneumoniae, here named MpgA and MpgB, are important in maintaining cell wall integrity. MpgA was predicted to be a lytic transglycosylase based on its homology to Escherichia coli MltG while the enzymatic activity of MpgB was unclear. Using nascent peptidoglycan substrates synthesized in vitro from the peptidoglycan precursor Lipid II, we report that both MpgA and MpgB are muramidases. We show that replacing a single amino acid in E. coli MltG with the corresponding amino acid from MpgA results in muramidase activity, allowing us to predict from the presence of this amino acid that other putative lytic transglycosylases actually function as muramidases. Strikingly, we report that MpgA and MpgB cut nascent peptidoglycan at different positions along the sugar backbone relative to the reducing end. MpgA produces much longer peptidoglycan oligomers and we show that its cleavage site selectivity is controlled by the LysM-like subdomain, which is also present in MltG. We propose that MltG’s ability to complement loss of MpgA in S. pneumoniae despite performing different cleavage chemistry is because it can cleave nascent peptidoglycan at the same distance from the lipid anchor.


2004 ◽  
Vol 48 (3) ◽  
pp. 728-738 ◽  
Author(s):  
Alexey Ruzin ◽  
Guy Singh ◽  
Anatoly Severin ◽  
Youjun Yang ◽  
Russell G. Dushin ◽  
...  

ABSTRACT The naturally occurring mannopeptimycins (formerly AC98-1 through AC98-5) are a novel class of glycopeptide antibiotics that are active against a wide variety of gram-positive bacteria. The structures of the mannopeptimycins suggested that they might act by targeting cell wall biosynthesis, similar to other known glycopeptide antibiotics; but the fact that the mannopeptimycins retain activity against vancomycin-resistant organisms suggested that they might have a unique mode of action. By using a radioactive mannopeptimycin derivative bearing a photoactivation ligand, it was shown that mannopeptimycins interact with the membrane-bound cell wall precursor lipid II [C55-MurNAc-(peptide)-GlcNAc] and that this interaction is different from the binding of other lipid II-binding antibiotics such as vancomycin and mersacidin. The antimicrobial activities of several mannopeptimycin derivatives correlated with their affinities toward lipid II, suggesting that the inhibition of cell wall biosynthesis was primarily through lipid II binding. In addition, it was shown that mannopeptimycins bind to lipoteichoic acid in a rather nonspecific interaction, which might facilitate the accumulation of antibiotic on the bacterial cell surface.


2007 ◽  
Vol 71 (4) ◽  
pp. 620-635 ◽  
Author(s):  
Jean van Heijenoort

SUMMARY This review is an attempt to bring together and critically evaluate the now-abundant but dispersed data concerning the lipid intermediates of the biosynthesis of bacterial peptidoglycan. Lipid I, lipid II, and their modified forms play a key role not only as the specific link between the intracellular synthesis of the peptidoglycan monomer unit and the extracytoplasmic polymerization reactions but also in the attachment of proteins to the bacterial cell wall and in the mechanisms of action of antibiotics with which they form specific complexes. The survey deals first with their detection, purification, structure, and preparation by chemical and enzymatic methods. The recent important advances in the study of transferases MraY and MurG, responsible for the formation of lipids I and II, are reported. Various modifications undergone by lipids I and II are described, especially those occurring in gram-positive organisms. The following section concerns the cellular location of the lipid intermediates and the translocation of lipid II across the cytoplasmic membrane. The great efforts made since 2000 in the study of the glycosyltransferases catalyzing the glycan chain formation with lipid II or analogues are analyzed in detail. Finally, examples of antibiotics forming complexes with the lipid intermediates are presented.


2021 ◽  
Vol 118 (36) ◽  
pp. e2103740118
Author(s):  
Atsushi Taguchi ◽  
Julia E. Page ◽  
Ho-Ching Tiffany Tsui ◽  
Malcolm E. Winkler ◽  
Suzanne Walker

The peptidoglycan cell wall is a macromolecular structure that encases bacteria and is essential for their survival. Proper assembly of the cell wall requires peptidoglycan synthases as well as membrane-bound cleavage enzymes that control where new peptidoglycan is made and inserted. Previous studies have shown that two membrane-bound proteins in Streptococcus pneumoniae, here named MpgA and MpgB, are important in maintaining cell wall integrity. MpgA was predicted to be a lytic transglycosylase based on its homology to Escherichia coli MltG, while the enzymatic activity of MpgB was unclear. Using nascent peptidoglycan substrates synthesized in vitro from the peptidoglycan precursor Lipid II, we report that both MpgA and MpgB are muramidases. We show that replacing a single amino acid in E. coli MltG with the corresponding amino acid from MpgA results in muramidase activity, allowing us to predict from the presence of this amino acid that other putative lytic transglycosylases actually function as muramidases. Strikingly, we report that MpgA and MpgB cut nascent peptidoglycan at different positions along the sugar backbone relative to the reducing end, with MpgA producing much longer peptidoglycan oligomers. We show that the cleavage site selectivity of MpgA is controlled by the LysM-like subdomain, which is required for its full functionality in cells. We propose that MltG’s ability to complement the loss of MpgA in S. pneumoniae despite performing different cleavage chemistry is because it can cleave nascent peptidoglycan at the same distance from the lipid anchor.


Author(s):  
Roy Skidmore

The long-necked secretory cells in Onchidoris muricata are distributed in the anterior sole of the foot. These cells are interspersed among ciliated columnar and conical cells as well as short-necked secretory gland cells. The long-necked cells contribute a significant amount of mucoid materials to the slime on which the nudibranch travels. The body of these cells is found in the subepidermal tissues. A long process extends across the basal lamina and in between cells of the epidermis to the surface of the foot. The secretory granules travel along the process and their contents are expelled by exocytosis at the foot surface.The contents of the cell body include the nucleus, some endoplasmic reticulum, and an extensive Golgi body with large numbers of secretory vesicles (Fig. 1). The secretory vesicles are membrane bound and contain a fibrillar matrix. At high magnification the similarity of the contents in the Golgi saccules and the secretory vesicles becomes apparent (Fig. 2).


2009 ◽  
Vol 27 (1) ◽  
pp. 65-92
Author(s):  
Susan Jones

This article explores the diversity of British literary responses to Diaghilev's project, emphasising the way in which the subject matter and methodologies of Diaghilev's modernism were sometimes unexpectedly echoed in expressions of contemporary British writing. These discussions emerge both in writing about Diaghilev's work, and, more discretely, when references to the Russian Ballet find their way into the creative writing of the period, serving to anchor the texts in a particular cultural milieu or to suggest contemporary aesthetic problems in the domain of literary aesthetics developing in the period. Figures from disparate fields, including literature, music and the visual arts, brought to their criticism of the Ballets Russes their individual perspectives on its aesthetics, helping to consolidate the sense of its importance in contributing to the inter-disciplinary flavour of modernism across the arts. In the field of literature, not only did British writers evaluate the Ballets Russes in terms of their own poetics, their relationship to experimentation in the novel and in drama, they developed an increasing sense of the company's place in dance history, its choreographic innovations offering material for wider discussions, opening up the potential for literary modernism's interest in impersonality and in the ‘unsayable’, discussions of the body, primitivism and gender.


2018 ◽  
Vol 27 (1) ◽  
pp. 5-21
Author(s):  
Amanda Dennis

Lying in ditches, tromping through mud, wedged in urns, trash bins, buried in earth, bodies in Beckett appear anything but capable of acting meaningfully on their environments. Bodies in Beckett seem, rather, synonymous with abjection, brokenness, and passivity—as if the human were overcome by its materiality: odours, pain, foot sores, decreased mobility. To the extent that Beckett's personae act, they act vaguely (wandering) or engage in quasi-obsessive, repetitive tasks: maniacal rocking, rotating sucking stones and biscuits, uttering words evacuated of sense, ceaseless pacing. Perhaps the most vivid dramatization of bodies compelled to meaningless, repetitive movement is Quad (1981), Beckett's ‘ballet’ for television, in which four bodies in hooded robes repeat their series ad infinitum. By 1981, has all possibility for intentional action in Beckett been foreclosed? Are we doomed, as Hamm puts it, to an eternal repetition of the same? (‘Moments for nothing, now as always, time was never and time is over, reckoning closed and story ended.’)This article proposes an alternative reading of bodily abjection, passivity and compulsivity in Beckett, a reading that implies a version of agency more capacious than voluntarism. Focusing on Quad as an illustrative case, I show how, if we shift our focus from the body's diminished possibilities for movement to the imbrication of Beckett's personae in environments (a mound of earth), things, and objects, a different story emerges: rather than dramatizing the impossibility of action, Beckett's work may sketch plans for a more ecological, post-human version of agency, a more collaborative mode of ‘acting’ that eases the divide between the human, the world of inanimate objects, and the earth.Movements such as new materialism and object-oriented ontology challenge hierarchies among subjects, objects and environments, questioning the rigid distinction between animate and inanimate, and the notion of the Anthropocene emphasizes the influence of human activity on social and geological space. A major theoretical challenge that arises from such discourses (including 20th-century challenges to the idea of an autonomous, willing, subject) is to arrive at an account of agency robust enough to survive if not the ‘death of the subject’ then its imbrication in the material and social environment it acts upon. Beckett's treatment of the human body suggests a version of agency that draws strength from a body's interaction with its environment, such that meaning is formed in the nexus between body and world. Using the example of Quad, I show how representations of the body in Beckett disturb the opposition between compulsivity (when a body is driven to move or speak in the absence of intention) and creative invention. In Quad, serial repetition works to create an interface between body and world that is receptive to meanings outside the control of a human will. Paradoxically, compulsive repetition in Beckett, despite its uncomfortable closeness to addiction, harnesses a loss of individual control that proposes a more versatile and ecologically mindful understanding of human action.


2019 ◽  
Author(s):  
Sylvia L. Rivera ◽  
Akbar Espaillat ◽  
Arjun K. Aditham ◽  
Peyton Shieh ◽  
Chris Muriel-Mundo ◽  
...  

Transpeptidation reinforces the structure of cell wall peptidoglycan, an extracellular heteropolymer that protects bacteria from osmotic lysis. The clinical success of transpeptidase-inhibiting β-lactam antibiotics illustrates the essentiality of these cross-linkages for cell wall integrity, but the presence of multiple, seemingly redundant transpeptidases in many bacterial species makes it challenging to determine cross-link function precisely. Here we present a technique to covalently link peptide strands by chemical rather than enzymatic reaction. We employ bio-compatible click chemistry to induce triazole formation between azido- and alkynyl-D-alanine residues that are metabolically installed in the cell walls of Gram-positive and Gram-negative bacteria. Synthetic triazole cross-links can be visualized by substituting azido-D-alanine with azidocoumarin-D-alanine, an amino acid derivative that undergoes fluorescent enhancement upon reaction with terminal alkynes. Cell wall stapling protects the model bacterium Escherichia coli from β-lactam treatment. Chemical control of cell wall structure in live bacteria can provide functional insights that are orthogonal to those obtained by genetics.<br>


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