title: Molecular interaction of ferredoxin and ferredoxin-NADP+ reductase from human malaria parasite.
abstract: The malaria parasite possesses plant-type ferredoxin (Fd) and ferredoxin-NADP(+) reductase (FNR) in a plastid-derived organelle called the apicoplast. This Fd/FNR redox system, which potentially provides reducing power for essential biosynthetic pathways in the apicoplast, has been proposed as a target for the development of specific new anti-malarial agents. We studied the molecular interaction of Fd and FNR of human malaria parasite (Plasmodium falciparum), which were produced as recombinant proteins in Escherichia coli. NMR chemical shift perturbation analysis mapped the location of the possible FNR interaction sites on the surface of P. falciparum Fd. Site-specific mutation of acidic Fd residues in these regions and the resulting analyses of electron transfer activity and affinity chromatography of those mutants revealed that two acidic regions (a region including Asp26, Glu29 and Glu34, and the other including Asp65 and Glu66) dominantly contribute to the electrostatic interaction with P. falciparum FNR. The combination of Asp26/Glu29/Glu34 conferred a larger contribution than that of Asp65/Glu66, and among Asp26, Glu29 and Glu34, Glu29 was shown to be the most important residue for the interaction with P. falciparum FNR. These findings provide the basis for understanding molecular recognition between Fd and FNR of the malaria parasite.
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Japan. a-yoko@protein.osaka-u.ac.jp
authors.forename: Yoko
authors.initials: Y
authors.lastname: Kimata-Ariga
authors.forename: Takashi
authors.initials: T
authors.lastname: Saitoh
authors.forename: Takahisa
authors.initials: T
authors.lastname: Ikegami
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Horii
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Hase
mesh_terms: D000818:Animals; D005287:Ferredoxin-NADP Reductase / Q000737:chemistry*; D005288:Ferredoxins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D019906:Nuclear Magnetic Resonance, Biomolecular; D010944:Plants / Q000201:enzymology; D010963:Plasmodium falciparum / Q000737:chemistry* / Q000201:enzymology; D015800:Protozoan Proteins / Q000737:chemistry*
chemical_list: D005288:Ferredoxins; D015800:Protozoan Proteins; D005287:Ferredoxin-NADP Reductase