Probing the Mycobacterial Trehalome with Bioorthogonal Chemistry

Swarts, Benjamin M.; Holsclaw, Cynthia M.; Jewett, John C.; Alber, Marina; Fox, Douglas; Siegrist, M. Sloan; Leary, Julie A.; Kalscheuer, Rainer; Bertozzi, Carolyn R.

doi:10.1021/ja3062419

Cited by 165 publications

(278 citation statements)

References 25 publications

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“…The enzyme has been biochemically characterized from M. smegmatis, but in an attempt to determine the role of trehalase for trehalose catabolism in this organism, the authors were unable to inactivate the respective gene, suggesting that it is essential (31). Our group, however, was able to delete the trehalase gene from M. smegmatis without noticing any negative effect on viability (32). According to its suggested role, the trehalase-deficient M. smegmatis mutant was unable to utilize trehalose as the sole carbon source, confirming its crucial role in intracellular trehalose breakdown (M. Alber and R. Kalscheuer, unpublished).…”

Section: Trehalose Degradationmentioning

confidence: 74%

Genetics of Mycobacterial Trehalose Metabolism

Kalscheuer

Koliwer‐Brandl

2014

Microbiol Spectr

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Trehalose [alpha-D-glucopyranosyl-(1→1)-alpha- D-glucopyranoside] is a highly abundant disaccharide in mycobacteria that fulfills many biological roles and has a plethora of possible metabolic fates. Trehalose is synthesized in mycobacteria de novo either from glycolytic intermediates or from alpha-glucans via two alternative routes, the OtsA-OtsB and the TreY-TreZ pathways, respectively. Intracellular trehalose can serve as an endogenous remobilizable carbon storage compound and as a biocompatible stress protectant. Furthermore, trehalose functions as the sugar core of many glycolipids with important structural or immunomodulatory functions such as the cord factor trehalose dimycolate, sulfolipids, and polyacyltrehalose. Moreover, trehalose plays a central role in the formation of the mycolic acid cell wall layer because it serves as a carrier molecule that shuttles mycolic acids in the form of the glycolipid trehalose monomycolate between the cytoplasm and the periplasm. In this process, a specific importer recycles the free trehalose that is extracellularly released as a by-product during mycolate processing via the antigen 85 complex, which might represent a specific adaptation to the intracellular lifestyle of Mycobacterium tuberculosis with limited carbohydrate availability. Finally, trehalose is converted to glycogen-like branched alpha-glucans by a four-step metabolic pathway involving the essential maltosyltransferase GlgE, which may be further processed to derivatives with intracellular or extracellular destinations such as polymethylated lipopolysaccharides or capsular alpha-glucans, respectively. In this article we summarize the current knowledge of the genetic basis of trehalose biosynthesis and metabolism in mycobacteria, the biological functions of trehalose-based molecules, and their roles in virulence of the human pathogen M. tuberculosis.

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Section: Trehalose Degradationmentioning

confidence: 74%

Genetics of Mycobacterial Trehalose Metabolism

Kalscheuer

Koliwer‐Brandl

2014

Microbiol Spectr

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“…100 Referred to as TreAz analogues, these chemical reporters were shown to be metabolically incorporated into surface-bound trehalose glycolipids in various mycobacterial species, including M. smegmatis , M. bovis , and M. tuberculosis . Both CuAAC and SPAAC were used to deliver fluorophores to TreAz-labeled cells and permit subsequent detection by flow cytometry and fluorescence microscopy (Figure 10B).…”

Section: Applications Of Trehalose Analoguesmentioning

confidence: 99%

“…The syntheses of 2- and 3-TreAz required 7–9 steps and proceeded in very low overall yield from trehalose. 4- and 6-TreAz were also accessed via reported trehalose desymmetrization methods, with the former taking 6 steps 100,102 from trehalose and the latter taking 4 steps. 103 Although these syntheses were successful and provided enough material for evaluation, they were inefficient in terms of step count, time, and yield.…”

Section: Applications Of Trehalose Analoguesmentioning

confidence: 99%

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Tailoring trehalose for biomedical and biotechnological applications

O'Neill

Piligian

Olson

et al. 2017

Pure and Applied Chemistry

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Trehalose is a non-reducing sugar whose ability to stabilize biomolecules has brought about its widespread use in biological preservation applications. Trehalose is also an essential metabolite in a number of pathogens, most significantly the global pathogen Mycobacterium tuberculosis, though it is absent in humans and other mammals. Recently, there has been a surge of interest in modifying the structure of trehalose to generate analogues that have applications in biomedical research and biotechnology. Non-degradable trehalose analogues could have a number of advantages as bioprotectants and food additives. Trehalose-based imaging probes and inhibitors are already useful as research tools and may have future value in the diagnosis and treatment of tuberculosis, among other uses. Underlying the advancements made in these areas are novel synthetic methods that facilitate access to and evaluation of trehalose analogues. In this review, we focus on both aspects of the development of this class of molecules. First, we consider the chemical and chemoenzymatic methods that have been used to prepare trehalose analogues and discuss their prospects for synthesis on commercially relevant scales. Second, we describe ongoing efforts to develop and deploy detectable trehalose analogues, trehalose-based inhibitors, and non-digestible trehalose analogues. The current and potential future uses of these compounds are discussed, with an emphasis on their roles in understanding and combatting mycobacterial infection.

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“…5a,b) 71 . Azide-functionalized trehalose analogs have been generated as bioorthogonal chemical reporters for imaging 72,73 . This strategy is suitable for studying mycobacterial glycolipid function and dynamics in living cells.…”

Section: Carbohydrate-derived Chemical Probesmentioning

confidence: 99%

Progress and prospects for small-molecule probes of bacterial imaging

Kocaoglu

Carlson

2016

Nat Chem Biol

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Fluorescence microscopy is an essential tool for the exploration of cell growth, division, transcription and translation in eukaryotes and prokaryotes alike. Despite the rapid development of techniques to study bacteria, the size of these organisms (1–10 μm) and their robust and largely impenetrable cell envelope present major challenges in imaging experiments. Fusion-based strategies, such as attachment of the protein of interest to a fluorescent protein or epitope tag, are by far the most common means for examining protein localization and expression in prokaryotes. While valuable, the use of genetically encoded tags can result in mislocalization or altered activity of the desired protein, does not provide a readout of the catalytic state of enzymes and cannot enable visualization of many other important cellular components, such as peptidoglycan, lipids, nucleic acids or glycans. Here, we highlight the use of biomolecule-specific small-molecule probes for imaging in bacteria.

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Probing the Mycobacterial Trehalome with Bioorthogonal Chemistry

Cited by 165 publications

References 25 publications

Genetics of Mycobacterial Trehalose Metabolism

Genetics of Mycobacterial Trehalose Metabolism

Tailoring trehalose for biomedical and biotechnological applications

Progress and prospects for small-molecule probes of bacterial imaging

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