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Biochemical characterization of Plasmodium falciparum CTP:phosphoethanolamine cytidylyltransferase shows that only one of the two cytidylyltransferase domains is active
- Source :
- Biochemical Journal; Vol 450, Biochemical Journal, Biochemical Journal, Portland Press, 2013, 450 (1), pp.159-167. ⟨10.1042/BJ20121480⟩
- Publication Year :
- 2013
- Publisher :
- PORTLAND PRESS LTD, 2013.
-
Abstract
- The intra-erythrocytic proliferation of the human malaria parasite Plasmodium falciparum requires massive synthesis of PE (phosphatidylethanolamine) that together with phosphatidylcholine constitute the bulk of the malaria membrane lipids. PE is mainly synthesized de novo by the CDP:ethanolamine-dependent Kennedy pathway. We previously showed that inhibition of PE biosynthesis led to parasite death. In the present study we characterized PfECT [P. falciparum CTP:phosphoethanolamine CT (cytidylyltransferase)], which we identified as the rate-limiting step of the PE metabolic pathway in the parasite. The cellular localization and expression of PfECT along the parasite life cycle were studied using polyclonal antibodies. Biochemical analyses showed that the enzyme activity follows Michaelis–Menten kinetics. PfECT is composed of two CT domains separated by a linker region. Activity assays on recombinant enzymes upon site-directed mutagenesis revealed that the N-terminal CT domain was the only catalytically active domain of PfECT. Concordantly, three-dimensional homology modelling of PfECT showed critical amino acid differences between the substrate-binding sites of the two CT domains. PfECT was predicted to fold as an intramolecular dimer suggesting that the inactive C-terminal domain is important for dimer stabilization. Given the absence of PE synthesis in red blood cells, PfECT represents a potential antimalarial target opening the way for a rational conception of bioactive compounds.
- Subjects :
- Models, Molecular
Protein Conformation
Membrane lipids
Cytidylyltransferase
Plasmodium falciparum
Protozoan Proteins
[CHIM.THER]Chemical Sciences/Medicinal Chemistry
Biology
Biochemistry
03 medical and health sciences
chemistry.chemical_compound
Mice
Biosynthesis
Animals
Humans
Molecular Biology
Cellular localization
ComputingMilieux_MISCELLANEOUS
030304 developmental biology
chemistry.chemical_classification
Phosphatidylethanolamine
0303 health sciences
Mice, Inbred BALB C
Binding Sites
Phosphatidylethanolamines
030302 biochemistry & molecular biology
RNA Nucleotidyltransferases
Cell Biology
[SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences
biology.organism_classification
3. Good health
Amino acid
Protein Structure, Tertiary
Kinetics
Enzyme
chemistry
Female
[INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM]
[CHIM.CHEM]Chemical Sciences/Cheminformatics
Subjects
Details
- Language :
- English
- ISSN :
- 14708728 and 02646021
- Volume :
- 450
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Biochemical Journal
- Accession number :
- edsair.doi.dedup.....b2f2421f677c8388b70e1efd93be985d
- Full Text :
- https://doi.org/10.1042/BJ20121480