Vitamin B6 biosynthesis by the malaria parasite Plasmodium falciparum: biochemical and structural insights

• Main Authors: Gengenbacher, Martin (Author), Fitzpatrick, Teresa B (Author), Raschle, Thomas (Author), Flicker, Karlheinz (Author), Sinning, Irmgard (Author), Müller, Sylke (Author), Macheroux, Peter (Author), Tews, Ivo (Author), Kappes, Barbara (Author)
• Format: Article
• Language: English
• Published: 2006-02-10.
• Subjects: Article
• Online Access: Get fulltext; Get fulltext

 Vitamin B6 is one of nature's most versatile cofactors. Most organisms synthesize vitamin B6 via a recently discovered pathway employing the proteins Pdx1 and Pdx2. Here we present an in-depth characterization of the respective orthologs from the malaria parasite, Plasmodium falciparum. Expression profiling of Pdx1 and -2 shows that blood-stage parasites indeed possess a functional vitamin B6 de novo biosynthesis. Recombinant Pdx1 and Pdx2 form a complex that functions as a glutamine amidotransferase with Pdx2 as the glutaminase and Pdx1 as pyridoxal-5 '-phosphate synthase domain. Complex formation is required for catalytic activity of either domain. Pdx1 forms a chimeric bi-enzyme with the bacterial YaaE, a Pdx2 ortholog, both in vivo and in vitro, although this chimera does not attain full catalytic activity, emphasizing that species-specific structural features govern the interaction between the protein partners of the PLP synthase complexes in different organisms. To gain insight into the activation mechanism of the parasite bi-enzyme complex, the three-dimensional structure of Pdx2 was determined at 1.62 A. The obstruction of the oxyanion hole indicates that Pdx2 is in a resting state and that activation occurs upon Pdx1-Pdx2 complex formation.