Engineering the Mycomembrane of Live Mycobacteria with an Expanded Set of Trehalose Monomycolate Analogues
| Autor: | Benjamin M. Swarts, Adrian K. Rylski, Taylor J. Fiolek, Herbert W. Kavunja, Nathan J. Holmes, Amol Arunrao Pohane, Nicholas Banahene, M. Sloan Siegrist |
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| Rok vydání: | 2019 |
| Předmět: |
Azides
Mycobacterium smegmatis Context (language use) Corynebacterium 010402 general chemistry 01 natural sciences Biochemistry Article Tetrazine chemistry.chemical_compound Biosynthesis Corynebacterineae Escherichia coli Cell Engineering Molecular Biology Fluorescent Dyes Molecular Structure biology 010405 organic chemistry Cell Membrane Organic Chemistry Mycobacterium tuberculosis biology.organism_classification 0104 chemical sciences chemistry Alkynes Click chemistry Cord Factors Molecular Medicine Click Chemistry Azide Bacterial outer membrane Acyltransferases Function (biology) Bacillus subtilis |
| Zdroj: | Chembiochem |
| ISSN: | 1439-7633 1439-4227 |
| DOI: | 10.1002/cbic.201800687 |
| Popis: | Mycobacteria and related organisms in the Corynebacterineae suborder are characterized by a distinctive outer membrane referred to as the mycomembrane. Biosynthesis of the mycomembrane occurs through an essential process called mycoloylation, which involves antigen 85 (Ag85)-catalyzed transfer of mycolic acids from the mycoloyl donor trehalose monomycolate (TMM) to acceptor carbohydrates and, in some organisms, proteins. We recently described an alkyne-modified TMM analogue (O-AlkTMM-C7) which, in conjunction with click chemistry, acted as a chemical reporter for mycoloylation in intact cells and allowed metabolic labeling of mycoloylated components of the mycomembrane. Here, we describe the synthesis and evaluation of a toolbox of TMM-based reporters bearing alkyne, azide, trans-cyclooctene, and fluorescent tags. These compounds gave further insight into the substrate tolerance of mycoloyltransferases (e.g., Ag85s) in a cellular context and they provide significantly expanded experimental versatility by allowing one- or two-step cell labeling, live cell labeling, and rapid cell labeling via tetrazine ligation. Such capabilities will facilitate research on mycomembrane composition, biosynthesis, and dynamics. Moreover, because TMM is exclusively metabolized by Corynebacterineae, the described probes may be valuable for the specific detection and cell-surface engineering of Mycobacterium tuberculosis and related pathogens. We also performed experiments to establish the dependence of probe incorporation on mycoloyltransferase activity, results from which suggested that cellular labeling is a function not only of metabolic incorporation (and likely removal) pathway(s), but also accessibility across the envelope. Thus, whole-cell labeling experiments with TMM reporters should be carefully designed and interpreted when envelope permeability may be compromised. On the other hand, this property of TMM reporters can potentially be exploited as a convenient way to probe changes in envelope integrity and permeability, facilitating drug development studies. |
| Databáze: | OpenAIRE |
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