PAN · Mechanism and lnhibition of pantothenate synthase

Lines connect the coordinator with its partners.

Project objective

Pantothenate is vitamin B5, an essential dietary vitamin for humans and animals. The biosyntheticpathway leading to pantothenate has been elucidated, and the enzymes and their correspondinggenes isolated from E. coli. Since pantothenate is made by microorg anisms, plants and fungi, theenzymes which form pantothenate are potentially excellent targets for the design of antimicrobial 01herbicida! agents. This project centers on the enzyme pantothenate synthase, which catalyses the finalstep in the formation of pantothenate. Pantothenate synthase is readily available throughoverexpression of the cloned gene. The recently determined crystal structure of this enzyme will beused as a guide for both parts of this project:(i) detailed studies of the mechanism of actio nSite-directed mutagenesis will be used to probe the mechanistic role of key residues identified fromthe crystal structur, and to form stable enzyme-substrate complexes for structural studies. Thesubstrate specificity of pantothenate synthase will be explo red and attempts made to alter thesubstrate specificity by mutagenesis.(ii) design of enzyme inhibitors.Novel strategies will be used for rational inhibitor design, including Click chemistry andenzyme-templated dynamic combinatorial chemistry (DCC). Studie s using Click chemistry will initiallyuse a range of azides and alkynes based on the pantoate and adenosine, reacting via templating inthe presence of the enzyme, and so favouring products which bind the enzyme most tightly. Enzyme-templated DCC will invol ve the reversible reactions of pools of molecular fragments using excessenzyme to template the formation of products which bind tightly to the enzyme and so are potentialinhibitors. Fragments for both Click and DCC studies will be chosen using state-of-the -art techniquesincluding in silico molecular docking using GOLD, small molecule-protein binding by 1H NMR.

Original text from CORDIS.

Participants

• University of Cambridge · CambridgeCoordinatorUnited Kingdom

Links

• View on CORDIS