tetrahydrodipicolinate n-succinyltransferase and dihydrodipicolinate synthase from pseudomonas aeruginosa structure analysis and gene deletiontetrahydrodipicolinate n-succinyltransferase dihydrodipicolinate合成酶和铜绿假单胞菌结构分析和基因删除.pdfVIP

Tetrahydrodipicolinate N-Succinyltransferase and Dihydrodipicolinate Synthase from Pseudomonas aeruginosa : Structure Analysis and Gene Deletion 1 2 1 2 3 Robert Schnell , Wulf Oehlmann , Tatyana Sandalova , Yvonne Braun , Carmen Huck , Marko 3 2 1 Maringer , Mahavir Singh *, Gunter Schneider * 1 Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden, 2 LIONEX Diagnostics and Therapeutics, Braunschweig, Germany, 3 mfd Diagnostics GmbH, Wendelsheim, Germany Abstract The diaminopimelic acid pathway of lysine biosynthesis has been suggested to provide attractive targets for the development of novel antibacterial drugs. Here we report the characterization of two enzymes from this pathway in the human pathogen Pseudomonas aeruginosa, utilizing structural biology, biochemistry and genetics. We show that tetrahydrodipicolinate N-succinyltransferase (DapD) from P. aeruginosa is specific for the L-stereoisomer of the amino substrate L-2-aminopimelate, and its D-enantiomer acts as a weak inhibitor. The crystal structures of this enzyme with L-2- aminopimelate and D-2-aminopimelate, respectively, reveal that both compounds bind at the same site of the enzyme. Comparison of the binding interactions of these ligands in the enzyme active site suggests misalignment of the amino group of D-2-aminopimelate for nucleophilic attack on the succinate moiety of the co-substrate succinyl-CoA as the structural basis of specificity and inhibition. P. aeruginosa mutants where the dapA gene had been deleted were viable and able t