tailieunhanh - Báo cáo khoa học: Characterization ofAquifex aeolicus4-diphosphocytidyl-2C-methyl-D-erythritol kinase – ligand recognition in a template for antimicrobial drug discovery

4-Diphosphocytidyl-2C-methyl-d-erythritol kinase (IspE) catalyses the ATP-dependent conversion of 4-diphosphocytidyl-2C-methyl-d-erythritol (CDPME) to 4-diphosphocytidyl-2C-methyl-d-erythritol 2-phosphate with the release of ADP. | ỊFEBS Journal Characterization of Aquifex aeolicus 4-diphosphocytidyl-2C-methyl-D-erythritol kinase - ligand recognition in a template for antimicrobial drug discovery Tanja Sgraja1 Magnus S. Alphey1 Stephanos Ghilagaber1 Rudi Marquez2 Murray N. Robertson2 Jennifer L. Hemmings2 Susan Lauw3 Felix Rohdich3 Adelbert Bacher3 Wolfgang Eisenreich3 Victoria Illarionova3 and William N. Hunter1 1 Division of BiologicalChemistry and Drug Discovery University of Dundee UK 2 Department of Chemistry University of Glasgow UK 3 Center for Integrated Protein Research Technische Universitat Munchen Garching Germany QnlineGpen Thisarticleis available free online at Keywords enzyme-ligand complex GHMP kinase isoprenoid biosynthesis molecular recognition non-mevalonate pathway Correspondence W. M. Hunter Division of Biological Chemistry and Drug Discovery College of Life Sciences University of Dundee Dundee DD1 5EH UK Fax 44 1382 385764 Tel 44 1382 385745 E-mail Re-use of this article is permitted in accordance with the Creative Commons Deed Attribution which does not permit commercial exploitation Received 4 February 2008 revised 13 March 2008 accepted 20 March 2008 doi 4-Diphosphocytidyl-2C-methyl-D-erythritol kinase IspE catalyses the ATP-dependent conversion of 4-diphosphocytidyl-2C-methyl-D-erythritol CDPME to 4-diphosphocytidyl-2C-methyl-D-erythritol 2-phosphate with the release of ADP. This reaction occurs in the non-mevalonate pathway of isoprenoid precursor biosynthesis and because it is essential in important microbial pathogens and absent from mammals it represents a potential target for anti-infective drugs. We set out to characterize the biochemical properties determinants of molecular recognition and reactivity of IspE and report the cloning and purification of recombinant Aquifex aeolicus IspE AaIspE kinetic data metal ion temperature and pH dependence crystallization and structure .