N α -acetyl-L-ornithine deacetylase from Escherichia coli and a ninhydrin-based assay to enable inhibitor identification
Emma H. Kelley, Jerzy Osipiuk, Malgorzata Korbas, Michael Endres, Alayna Bland, Victoria Ehrman, Andrzej Joachimiak, Kenneth W. Olsen, Daniel P. Becker
TL;DR
Researchers developed a new assay to identify inhibitors of a bacterial enzyme, ArgE, and found captopril and phenylboronic acids as potential antibiotic candidates.
Contribution
A novel ninhydrin-based assay for ArgE was developed, enabling inhibitor screening and leading to the discovery of new inhibitors.
Findings
Captopril inhibits ArgE with an IC50 of 58.7 μM and Ki of 37.1 ± 0.85 μM.
Phenylboronic acid derivatives, including 4-(diethylamino)phenylboronic acid, were identified as ArgE inhibitors.
X-ray crystal structures of E. coli ArgE (mono-Zn and di-Zn) were reported for the first time.
Abstract
Bacteria are becoming increasingly resistant to antibiotics, therefore there is an urgent need for new classes of antibiotics to fight antibiotic resistance. Mammals do not express N ɑ -acetyl-L-ornithine deacetylase (ArgE), an enzyme that is critical for bacterial survival and growth, thus ArgE represents a promising new antibiotic drug target, as inhibitors would not suffer from mechanism-based toxicity. A new ninhydrin-based assay was designed and validated that included the synthesis of the substrate analog N 5, N 5-di-methyl N α-acetyl-L-ornithine (kcat/Km = 7.32 ± 0.94 × 104 M−1s−1). This new assay enabled the screening of potential inhibitors that absorb in the UV region, and thus is superior to the established 214 nm assay. Using this new ninhydrin-based assay, captopril was confirmed as an ArgE inhibitor (IC50 = 58.7 μM; Ki = 37.1 ± 0.85 μM), and a number of phenylboronic acid…
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Taxonomy
TopicsPolyamine Metabolism and Applications · Enzyme Structure and Function · Amino Acid Enzymes and Metabolism