Plasmodium falciparum CTP:phosphocholine cytidylyltransferase possesses two functional catalytic domains and is inhibited by a CDP-choline analog selected from a virtual screening

FEBS Lett. 2015 Apr 13;589(9):992-1000. doi: 10.1016/j.febslet.2015.03.003. Epub 2015 Mar 13.

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.

Keywords: CTP:phosphocholine cytidylyltransferase (EC 2.7.7.15); Drug target; Inhibitor; Kinetic parameter; Malaria; Phosphatidylcholine; Structure-based virtual screening.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antimalarials / chemistry
  • Antimalarials / pharmacology
  • Biosynthetic Pathways / genetics
  • Catalytic Domain*
  • Choline-Phosphate Cytidylyltransferase / antagonists & inhibitors
  • Choline-Phosphate Cytidylyltransferase / genetics
  • Choline-Phosphate Cytidylyltransferase / metabolism*
  • Cytidine Diphosphate Choline / analogs & derivatives*
  • Cytidine Diphosphate Choline / chemistry
  • Cytidine Diphosphate Choline / pharmacology
  • Humans
  • Immunoblotting
  • Kinetics
  • Microscopy, Fluorescence
  • Models, Molecular
  • Molecular Sequence Data
  • Molecular Structure
  • Phosphatidylcholines / biosynthesis
  • Phosphatidylcholines / chemistry
  • Plasmodium falciparum / enzymology*
  • Plasmodium falciparum / genetics
  • Protein Binding
  • Protein Structure, Tertiary
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / genetics
  • Protozoan Proteins / metabolism*
  • Sequence Homology, Amino Acid

Substances

  • Antimalarials
  • Phosphatidylcholines
  • Protozoan Proteins
  • Cytidine Diphosphate Choline
  • Choline-Phosphate Cytidylyltransferase