tailieunhanh - Báo cáo khoa học: Phosphopantetheinyl transferase inhibition and secondary metabolism

Efforts to isolate carrier protein-mediated synthases from natural product-producing organisms using reporter-linked post-translational modification have been complicated by the efficiency of the endogenous process. To address this issue, we chose to target endogenous phosphopantetheinyl transferases (PPTases) for inhibitor design to facilitate natural product syn-thase isolation through a chemical genetics approach. | Phosphopantetheinyl transferase inhibition and secondary metabolism Timothy L. Foley Brian S. Young and Michael D. Burkart Department of Chemistry Biochemistry University of California San Diego CA USA Keywords enzyme inhibition fatty acid nonribosomal peptide phosphopantetheine polyketide Correspondence M. D. Burkart Department of Chemistry Biochemistry University of California San Diego 9500 Gilman Drive La Jolla CA 92093-0358 USA Fax 1 858 822 2182 Tel 1 858 534 5673 E-mail mburkart@ Received 27 July 2009 revised 16 September 2009 accepted 5 October 2009 doi Efforts to isolate carrier protein-mediated synthases from natural productproducing organisms using reporter-linked post-translational modification have been complicated by the efficiency of the endogenous process. To address this issue we chose to target endogenous phosphopantetheinyl transferases PPTases for inhibitor design to facilitate natural product synthase isolation through a chemical genetics approach. Herein we validate secondary metabolism-associated PPTase for chemical probe development. We synthesized and evaluated a panel of compounds based on the anthranilate 4H-oxazol-5-one pharmacophore previously described to attenuate PPTase activity within bacterial cultures. Through the use of a new high-throughput Forster resonance energy transfer assay we demonstrated that these compounds exclusively inhibit fatty acid synthase-specific PPTases. In vivo a lead compound within this panel demonstrated selective antibiotic activity in a Bacillus subtilis model. Further evaluation demonstrated that the compound enhances actinorhodin production in Streptomy-ces coelicolor revealing the ability of this class of molecules to stimulate precocious secondary metabolite production. Introduction Fatty acids nonribosomal peptides and polyketides represent three classes of metabolites that play important roles in human health disease and therapy 1-4 . Current studies of the