@article {2620,
	title = {Engineering the Mycomembrane of Live Mycobacteria with an Expanded Set of Trehalose Monomycolate Analogues.},
	journal = {Chembiochem},
	volume = {20},
	year = {2019},
	month = {2019 05 15},
	pages = {1282-1291},
	abstract = {<p>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.</p>
},
	keywords = {Acyltransferases, Alkynes, Azides, Bacillus subtilis, Cell Engineering, Cell Membrane, Click Chemistry, Cord Factors, Corynebacterium, Escherichia coli, Fluorescent Dyes, Molecular Structure, Mycobacterium smegmatis, Mycobacterium tuberculosis},
	issn = {1439-7633},
	doi = {10.1002/cbic.201800687},
	author = {Fiolek, Taylor J and Banahene, Nicholas and Kavunja, Herbert W and Holmes, Nathan J and Rylski, Adrian K and Pohane, Amol Arunrao and Siegrist, M Sloan and Swarts, Benjamin M}
}
@article {1445,
	title = {(D)-Amino acid chemical reporters reveal peptidoglycan dynamics of an intracellular pathogen.},
	journal = {ACS Chem Biol},
	volume = {8},
	year = {2013},
	month = {2013 Mar 15},
	pages = {500-5},
	abstract = {<p>Peptidoglycan (PG) is an essential component of the bacterial cell wall. Although experiments with organisms in vitro have yielded a wealth of information on PG synthesis and maturation, it is unclear how these studies translate to bacteria replicating within host cells. We report a chemical approach for probing PG in vivo via metabolic labeling and bioorthogonal chemistry. A wide variety of bacterial species incorporated azide and alkyne-functionalized d-alanine into their cell walls, which we visualized by covalent reaction with click chemistry probes. The d-alanine analogues were specifically incorporated into nascent PG of the intracellular pathogen Listeria monocytogenes both in vitro and during macrophage infection. Metabolic incorporation of d-alanine derivatives and click chemistry detection constitute a facile, modular platform that facilitates unprecedented spatial and temporal resolution of PG dynamics in vivo.</p>},
	keywords = {Alanine, Click Chemistry, Listeria monocytogenes, Molecular Conformation, Molecular Dynamics Simulation, Peptidoglycan},
	issn = {1554-8937},
	doi = {10.1021/cb3004995},
	author = {Siegrist, M Sloan and Whiteside, Sarah and Jewett, John C and Aditham, Arjun and Cava, Felipe and Bertozzi, Carolyn R}
}
