title: Plasmodium falciparum CTP:phosphocholine cytidylyltransferase possesses two functional catalytic domains and is inhibited by a CDP-choline analog selected from a virtual screening.
abstract: Phosphatidylcholine is the major lipid component of the malaria parasite membranes and is required for parasite multiplication in human erythrocytes. Plasmodium falciparum CTP:phosphocholine cytidylyltransferase (PfCCT) is the rate-limiting enzyme of the phosphatidylcholine biosynthesis pathway and thus considered as a potential antimalarial target. In contrast to its mammalian orthologs, PfCCT contains a duplicated catalytic domain. Here, we show that both domains are catalytically active with similar kinetic parameters. A virtual screening strategy allowed the identification of a drug-size molecule competitively inhibiting the enzyme. This compound also prevented phosphatidylcholine biosynthesis in parasites and exerted an antimalarial effect. This study constitutes the first step towards a rationalized design of future new antimalarial agents targeting PfCCT.
authors.affiliation: Université Montpellier, CNRS, Dynamique des Interactions Membranaires Normales et Pathologiques, UMR 5235, Place Eugène Bataillon, 34095 Montpellier, France.
authors.forename: Alicia
authors.initials: A
authors.lastname: Contet
authors.affiliation: CNRS, Université Nice Sophia-Antipolis, Institut de Pharmacologie Moléculaire et Cellulaire, UMR 7275, 660, route des Lucioles, Sophia Antipolis, 06560 Valbonne, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Pihan
authors.affiliation: Université Montpellier, CNRS, Dynamique des Interactions Membranaires Normales et Pathologiques, UMR 5235, Place Eugène Bataillon, 34095 Montpellier, France.
authors.forename: Marina
authors.initials: M
authors.lastname: Lavigne
authors.affiliation: Université Montpellier, CNRS, Dynamique des Interactions Membranaires Normales et Pathologiques, UMR 5235, Place Eugène Bataillon, 34095 Montpellier, France.
authors.forename: Kai
authors.initials: K
authors.lastname: Wengelnik
authors.affiliation: Université Montpellier, CNRS, Dynamique des Interactions Membranaires Normales et Pathologiques, UMR 5235, Place Eugène Bataillon, 34095 Montpellier, France.
authors.forename: Sweta
authors.initials: S
authors.lastname: Maheshwari
authors.affiliation: Université Montpellier, CNRS, Dynamique des Interactions Membranaires Normales et Pathologiques, UMR 5235, Place Eugène Bataillon, 34095 Montpellier, France.
authors.forename: Henri
authors.initials: H
authors.lastname: Vial
authors.affiliation: CNRS, Université Nice Sophia-Antipolis, Institut de Pharmacologie Moléculaire et Cellulaire, UMR 7275, 660, route des Lucioles, Sophia Antipolis, 06560 Valbonne, France. Electronic address: douguet@ipmc.cnrs.fr.
authors.forename: Dominique
authors.initials: D
authors.lastname: Douguet
authors.affiliation: Université Montpellier, CNRS, Dynamique des Interactions Membranaires Normales et Pathologiques, UMR 5235, Place Eugène Bataillon, 34095 Montpellier, France. Electronic address: rachel.cerdan@univ-montp2.fr.
authors.forename: Rachel
authors.initials: R
authors.lastname: Cerdan
mesh_terms: D000595:Amino Acid Sequence; D000962:Antimalarials / Q000737:chemistry / Q000494:pharmacology; D053898:Biosynthetic Pathways / Q000235:genetics; D020134:Catalytic Domain; D019916:Choline-Phosphate Cytidylyltransferase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D003566:Cytidine Diphosphate Choline / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D007700:Kinetics; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010713:Phosphatidylcholines / Q000096:biosynthesis / Q000737:chemistry; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D000962:Antimalarials; D010713:Phosphatidylcholines; D015800:Protozoan Proteins; D003566:Cytidine Diphosphate Choline; D019916:Choline-Phosphate Cytidylyltransferase
keywords: CTP:phosphocholine cytidylyltransferase (EC 2.7.7.15); Drug target; Inhibitor; Kinetic parameter; Malaria; Phosphatidylcholine; Structure-based virtual screening