1 Crystal structure of the Brucella abortus deoxyxylulose 5-phosphate reductoisomerase-like (DRL) enzyme involved in isoprenoid biosynthesis. Jordi Pérez-Gil* ,1 , Bárbara M. Calisto* ,2 , Christoph Behrendt 3 , Thomas Kurz 3 , Ignacio Fita 2 , and Manuel Rodríguez-Concepción 1 1, Department of Molecular Genetics, Centre for Research in Agricultural Genomics (CRAG) CSIC- IRTA-UAB-UB, Campus UAB Bellaterra, 08193 Barcelona, Spain. 2, Institut de Biologia Molecular de Barcelona (IBMB-CSIC) and Institute for Research in Biomedicine (IRB Barcelona), Parc Científic de Barcelona, Baldiri Reixac 10, 08028 Barcelona, Spain 3, Institut für Pharmazeustische und Medizinische Chemie, Heinrich-Heine-Universität Düsseldorf, Universitätsstr. 1, 40225 Düsseldorf, Germany * These authors contributed equally to the work. Running Title: Targeting Isoprenoid Enzymes for Novel Antibiotics To whom correspondence should be addressed: Manuel Rodríguez-Concepción, Department of Molecular Genetics, Centre for Research in Agricultural Genomics (CRAG), Campus UAB Bellaterra, 08193 Barcelona, Spain. Tel.: +34 935636600 ext 3222; Email: manuel.rodriguez@cragenomica.es Keywords: antibiotics; Brucella; isoprenoid, MEP pathway _____________________________________________________________________________________ Background: The antibiotic resistance epidemic demands new drugs specifically targeting the infective agents. Results: Crystal structure of the Brucella DRL enzyme shows major differences with DXR, which catalyzes the same reaction in most other bacteria. Conclusion: Structural information will allow development of inhibitors targeting only DRL. Significance: Drugs against DRL could function as highly-specific, narrow-range antibiotics. SUMMARY Most bacteria use the 2-C-methyl-D- erythritol 4-phosphate (MEP) pathway for the synthesis of their essential isoprenoid precursors. The absence of the MEP pathway in humans makes it a promising new target for the development of much needed new and safe antimicrobial drugs. However, bacteria show a remarkable metabolic plasticity for isoprenoid production. For example, the NADPH- dependent production of MEP from 1-deoxy- D-xylulose 5-phosphate (DXP) in the first committed step of the MEP pathway is catalyzed by DXP reductoisomerase (DXR) in most bacteria, whereas an unrelated DXR-like (DRL) protein was recently found to catalyze the same reaction in some organisms, including the emerging human and animal pathogens Bartonella and Brucella. Here we report the X- ray crystal structures of the Brucella abortus DRL enzyme in its apo form and in complex with the broad spectrum antibiotic fosmidomycin solved to 1.5 and 1.8 Å resolution, respectively. DRL is a dimer, with each polypeptide folding into three distinct domains starting with the NADPH-binding domain in resemblance to the structure of bacterial DXRs. Other than that, DRL and DXR show a low structural relationship with a different disposition of the domains and a topologically unrelated C-terminal domain. In http://www.jbc.org/cgi/doi/10.1074/jbc.M112.354811 The latest version is at JBC Papers in Press. Published on March 22, 2012 as Manuscript M112.354811 Copyright 2012 by The American Society for Biochemistry and Molecular Biology, Inc. by guest on February 12, 2016 http://www.jbc.org/ Downloaded from by guest on February 12, 2016 http://www.jbc.org/ Downloaded from by guest on February 12, 2016 http://www.jbc.org/ Downloaded from by guest on February 12, 2016 http://www.jbc.org/ Downloaded from by guest on February 12, 2016 http://www.jbc.org/ Downloaded from by guest on February 12, 2016 http://www.jbc.org/ Downloaded from by guest on February 12, 2016 http://www.jbc.org/ Downloaded from by guest on February 12, 2016 http://www.jbc.org/ Downloaded from by guest on February 12, 2016 http://www.jbc.org/ Downloaded from