2226-71-3Relevant articles and documents
Optimization of CoaD Inhibitors against Gram-Negative Organisms through Targeted Metabolomics
Rath, Christopher M.,Benton, Bret M.,De Vicente, Javier,Drumm, Joseph E.,Geng, Mei,Li, Cindy,Moreau, Robert J.,Shen, Xiaoyu,Skepper, Colin K.,Steffek, Micah,Takeoka, Kenneth,Wang, Lisha,Wei, Jun-Rong,Xu, Wenjian,Zhang, Qiong,Feng, Brian Y.
, p. 391 - 402 (2018/03/21)
Drug-resistant Gram-negative bacteria are of increasing concern worldwide. Novel antibiotics are needed, but their development is complicated by the requirement to simultaneously optimize molecules for target affinity and cellular potency, which can result in divergent structure-activity relationships (SARs). These challenges were exemplified during our attempts to optimize inhibitors of the bacterial enzyme CoaD originally identified through a biochemical screen. To facilitate lead optimization, we developed mass spectroscopy assays based on the hypothesis that levels of CoA metabolites would reflect the cellular enzymatic activity of CoaD. Using these methods, we were able to monitor the effects of cellular enzyme inhibition at compound concentrations up to 100-fold below the minimum inhibitory concentration (MIC), a common metric of growth inhibition. Furthermore, we generated a panel of efflux pump mutants to dissect the susceptibility of a representative CoaD inhibitor to efflux. These approaches allowed for a nuanced understanding of the permeability and efflux liabilities of the series and helped guide optimization efforts to achieve measurable MICs against wild-type E. coli.
One-pot chemo-enzymatic synthesis of reporter-modified proteins
Worthington, Andrew S.,Burkart, Michael D.
, p. 44 - 46 (2007/10/03)
To meet recent advancements in the covalent reporter labeling of proteins, we propose a flexible synthesis for reporter analogs. Here we demonstrate a one-pot chemo-enzymatic synthesis of reporter-labeled proteins that allows the covalent tethering of any amine-terminal fluorescent or affinity label to a carrier protein or fusion construct. This two-reaction sequence consists of activated panthothenate coupling, biosynthetic conversion to the coenzyme A (CoA) analog, and enzymatic carrier protein modification via phosphopantetheinyltransferase (PPTase). We also probe substrate specificity for CoAA, the first enzyme in the pathway. With this approach CoA analogs may be rapidly prepared, thus permitting the regiospecific attachment of reporter moieties from a variety of molecular species. The Royal Society of Chemistry 2006.
