the assembly of the plasmodial plp synthase complex follows a defined course的会中plasmodial plp合酶复杂之前定义的过程.pdfVIP

The Assembly of the Plasmodial PLP Synthase Complex Follows a Defined Course ¨ 1. ¨ 1. 2 2 3 Ingrid B. Muller , Julia Knockel , Matthew R. Groves , Rositsa Jordanova , Steven E. Ealick , Rolf D. 1 1 Walter , Carsten Wrenger * 1 Department of Biochemistry, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany, 2 European Molecular Biology Laboratory-Hamburg Outstation, Hamburg, Germany, 3 Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York, United States of America Abstract Background: Plants, fungi, bacteria and the apicomplexan parasite Plasmodium falciparum are able to synthesize vitamin B6 de novo, whereas mammals depend upon the uptake of this essential nutrient from their diet. The active form of vitamin B6 is pyridoxal 5-phosphate (PLP). For its synthesis two enzymes, Pdx1 and Pdx2, act together, forming a multimeric complex consisting of 12 Pdx1 and 12 Pdx2 protomers. Methodology/Principal Findings: Here we report amino acid residues responsible for stabilization of the structural and enzymatic integrity of the plasmodial PLP synthase, identified by using distinct mutational analysis and biochemical approaches. Residues R85, H88 and E91 (RHE) are located at the Pdx1:Pdx1 interface and play an important role in Pdx1 complex assembly. Mutation of these residues to alanine impedes both Pdx1 activity and Pdx2 binding. Furthermore, changing D26, K83 and K151 (DKK), amino acids from the active site of Pdx1, to alanine obstructs not only enzyme activity but also formation of the complex. In contrast to the monomeric appearance of the RHE mutant, alteration of the DKK