title: Localisation of Plasmodium falciparum uroporphyrinogen III decarboxylase of the heme-biosynthetic pathway in the apicoplast and characterisation of its catalytic properties.
abstract: Uroporphyrinogen decarboxylase (UROD) is a key enzyme in the heme-biosynthetic pathway and in Plasmodium falciparum it occupies a strategic position in the proposed hybrid pathway for heme biosynthesis involving shuttling of intermediates between different subcellular compartments in the parasite. In the present study, we demonstrate that an N-terminally truncated recombinant P. falciparum UROD (r(Delta)PfUROD) over-expressed and purified from Escherichia coli cells, as well as the native enzyme from the parasite were catalytically less efficient compared with the host enzyme, although they were similar in other enzyme parameters. Molecular modeling of PfUROD based on the known crystal structure of the human enzyme indicated that the protein manifests a distorted triose phosphate isomerase (TIM) barrel fold which is conserved in all the known structures of UROD. The parasite enzyme shares all the conserved or invariant amino acid residues at the active and substrate binding sites, but is rich in lysine residues compared with the host enzyme. Mutation of specific lysine residues corresponding to residues at the dimer interface in human UROD enhanced the catalytic efficiency of the enzyme and dimer stability indicating that the lysine rich nature and weak dimer interface of the wild-type PfUROD could be responsible for its low catalytic efficiency. PfUROD was localised to the apicoplast, indicating the requirement of additional mechanisms for transport of the product coproporphyrinogen to other subcellular sites for its further conversion and ultimate heme formation.
authors.affiliation: Department of Biochemistry, Indian Institute of Science, C.V. Raman Avenue, Bangalore, Karnataka 560 012, India.
authors.forename: Viswanathan Arun
authors.initials: VA
authors.lastname: Nagaraj
authors.forename: Rajavel
authors.initials: R
authors.lastname: Arumugam
authors.forename: Nagasuma R
authors.initials: NR
authors.lastname: Chandra
authors.forename: Dasari
authors.initials: D
authors.lastname: Prasad
authors.forename: Pundi N
authors.initials: PN
authors.lastname: Rangarajan
authors.forename: Govindarajan
authors.initials: G
authors.lastname: Padmanaban
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002384:Catalysis; D006418:Heme / Q000096:biosynthesis*; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010963:Plasmodium falciparum / Q000201:enzymology*; D011994:Recombinant Proteins / Q000235:genetics; D020539:Sequence Analysis, Protein; D015398:Signal Transduction / Q000235:genetics; D013045:Species Specificity; D014575:Uroporphyrinogen Decarboxylase / Q000235:genetics / Q000502:physiology*
chemical_list: D011994:Recombinant Proteins; D006418:Heme; D014575:Uroporphyrinogen Decarboxylase