Objective * To study the structure and kinetics of phosphofructokinase (PFK) and pyruvate kinase (PYK) of Leishmania, key enzymes in the metabolism of the parasite, and to determine differences with the corresponding mammalian enzymes.* To design and synthesize selective inhibitors of the Leishmania enzymes, based on their differences with the mammallian enzymes.Expected Outcome* The primary structure of Leishmania PFK and PYK will become available.* The enzymes will be overexpressed in a heterologous system and purified.* Kinetic parameters of the enzymes will be established.* Preliminary information on the structure of the enzymes will become available.* Compounds with (some) inhibitory activity on the Leishmania enzymes will be synthesized.* Cloning and sequence determination of the Leishmania PFK and PYK genes.* Overexpression of the Leishmania enzymes in bacteria (Escherichia coli) or yeast (Hansenula polymorpha).* Purification of the recombinant enzymes.* Kinetic analysis of the purified enzymes.* Structure modelling of the Leishmania enzymes, using the X-ray coordinates of the crystal structures of homologous enzymes.* Structure-function analysis of residues potentially important for inhibitor design by site-directed mutagenesis.* Crystallization trials of recombinant Leishmania PFK and PYK.* Synthesis of potentially selective inhibitors of Leishmania PFK and PYK. Fields of science engineering and technologymaterials engineeringcrystalsnatural sciencesbiological sciencesmicrobiologybacteriologynatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes Programme(s) FP4-INCO - Specific research, technological development and demonstration programme in the field of cooperation with third countries and international organizations, 1994-1998 Topic(s) 03010303 - Research on the biology preclinical models of diseases Call for proposal Data not available Funding Scheme CSC - Cost-sharing contracts Coordinator CHRISTIAN DE DUVE INSTITUTE OF CELLULAR PATHOLOGY EU contribution No data Address 74,Avenue Hippocrate 74, ICP-TROP 74.39 1200 BRUXELLES Belgium See on map Total cost No data Participants (3) Sort alphabetically Sort by EU Contribution Expand all Collapse all Universidad Central de Venezuela Venezuela EU contribution No data Address Calle Suapure 1020 Caracas See on map Total cost No data University of Edinburgh United Kingdom EU contribution No data Address George Square EH8 9XD Edinburgh See on map Total cost No data Université Paul Sabatier de Toulouse III France EU contribution No data Address 118,Route de Narbonne 31062 Toulouse See on map Total cost No data
