Deuterated carbohydrate probes as ‘label-free’ substrates for probing nutrient uptake in mycobacteria by nuclear reaction analysis

Lowery, Richard; Gibson, Matthew I.; Thompson, Richard C.; Fullam, Elizabeth

doi:10.1039/c4cc09588j

Cited by 9 publications

(11 citation statements)

References 16 publications

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“…Next, we probed whether modified trehalose analogs known to be imported by mycobacteria, including 2 H-trehalose, 2-azido-2-deoxy-α,α′-trehalose (2-azido-trehalose), 4-azido-4-deoxy-α,α′-trehalose (4-azido-trehalose), 6-azido-6-deoxy-α,α′-trehalose (6-azido-trehalose), α-D-mannopyranosyl-(1→1)-α-D-glucopyranoside (mannotrehalose), and α-D-galactopyranosyl-(1→1)-α-D-glucopyranoside (galactotrehalose) (see SI methods for synthetic details), are substrates of Mtr LpqY ( 11 , 12 , 13 , 14 ). All of these disaccharides influenced the melting temperature of Mtr LpqY but to a lesser extent ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Structural basis of trehalose recognition by the mycobacterial LpqY-SugABC transporter

Furze

Delso

Casal

et al. 2021

Journal of Biological Chemistry

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The Mycobacterium tuberculosis ( Mtb ) LpqY-SugABC ATP-binding cassette transporter is a recycling system that imports trehalose released during remodeling of the Mtb cell-envelope. As this process is essential for the virulence of the Mtb pathogen, it may represent an important target for tuberculosis drug and diagnostic development, but the transporter specificity and molecular determinants of substrate recognition are unknown. To address this, we have determined the structural and biochemical basis of how mycobacteria transport trehalose using a combination of crystallography, saturation transfer difference NMR, molecular dynamics, site-directed mutagenesis, biochemical/biophysical assays, and the synthesis of trehalose analogs. This analysis pinpoints key residues of the LpqY substrate binding lipoprotein that dictate substrate-specific recognition and has revealed which disaccharide modifications are tolerated. These findings provide critical insights into how the essential Mtb LpqY-SugABC transporter reuses trehalose and modified analogs and specifies a framework that can be exploited for the design of new antitubercular agents and/or diagnostic tools.

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Section: Resultsmentioning

confidence: 99%

Structural basis of trehalose recognition by the mycobacterial LpqY-SugABC transporter

Furze

Delso

Casal

et al. 2021

Journal of Biological Chemistry

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“…We have taken advantage of these facts in a proof-of-concept study on how to identify ABC transporters for a given substrate through lipoproteomic analyses. To date, the identification of ABC transporters and their respective substrates has mainly been based on in silico analysis followed by uptake assays using isotope-labeled probes (Lowery et al, 2015;Titgemeyer et al, 2007), binding assays in combination with X-ray analysis using purified SBPs and predicted ligands (Fullam et al, 2016;Jiang et al, 2014), or screening of transposon libraries and subsequent genome sequencing, searching for clones that are unable to grow on the substrate of interest (Braibant et al, 2000;Gopinath et al, 2013;Nazarova et al, 2017). This concept requires the long and tedious process of mutant library preparation and is not necessarily restricted to identifying uptake pathways, but also responds to metabolic genes.…”

Section: Introductionmentioning

confidence: 99%

Detection and Characterization of a Mycobacterial L-Arabinofuranose ABC Transporter Identified with a Rapid Lipoproteomics Protocol

Müller

Fröhlich

et al. 2019

Cell Chemical Biology

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“…93 Recently, 2 H-labeled trehalose was chemically synthesized through a one-step ruthenium-catalyzed deuteration of trehalose, and the product was used in conjunction with nuclear reaction analysis to evaluate trehalose uptake by M. smegmatis . 94 …”

Section: Applications Of Trehalose Analoguesmentioning

confidence: 99%

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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Deuterated carbohydrate probes as ‘label-free’ substrates for probing nutrient uptake in mycobacteria by nuclear reaction analysis

Abstract: Deuterated sugars that are transported into mycobacteria can be detected using ion beam (nuclear reaction) analysis.

Cited by 9 publications

References 16 publications

Structural basis of trehalose recognition by the mycobacterial LpqY-SugABC transporter

Structural basis of trehalose recognition by the mycobacterial LpqY-SugABC transporter

Detection and Characterization of a Mycobacterial L-Arabinofuranose ABC Transporter Identified with a Rapid Lipoproteomics Protocol

Tailoring trehalose for biomedical and biotechnological applications

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