1H, 13C, and 15N chemical shift assignments for PfPMT, a phosphoethanolamine methyltransferase from Plasmodium falciparum

Abstract

Phosphoethanolamine methyltransferases (PMTs also known as PEAMTs) catalyze the three-step s-adenosylmethionione-dependent methylation of phosphoethanolamine to form phosphocholine. These enzymes play an important function in the synthesis of phosphatidylcholine, the major phospholipid in the membranes of lower and higher eukaryotes, as well as in the production of the compatible solute and osmoprotectant glycine betaine in plants. Genetic studies in plants, Caenhorhabditis elegans and Plasmodium falciparum have demonstrated that disruption of PMT activity results in severe defects in important cellular processes such as development, replication, survival and sexual maturation and differentiation. Here we report chemical shift assignments for PfPMT, the PMT from Plasmodium falciparum. X-ray crystal structures have been recently reported for complexes of PfPMT, but the structure of the apoenzyme remains unknown. The solution structure of the apoenzyme will help to elucidate important details of the mechanism of substrate binding by PfPMT, as residues comprising the substrate binding site are inaccessible to solvent in the conformation evident in the available crystal structures. In addition to enabling determination of the solution structure of the apoenzyme, the assignments will facilitate additional investigations into the interaction of PfPMT with its substrates and inhibitors.

Role of phospholipid synthesis in the development and differentiation of malaria parasites in the blood

“…(49). The 3D structure of PfPMT and subsequently that of PvPMT (P. vivax phosphoethanolamine methyltransferase) and PkPMT (P. knowlesi phosphoethanolamine methyltransferase) showed high conservation, functional activity, and inhibition by the same drugs (29,52,53). Like other methyltransferases known to be inhibited by the 4-aminoquinoline, amodiaquine, PfPMT was also inhibited by this compound (Table 2) (54).…”

Characterization of Plasmodium phosphatidylserine decarboxylase expressed in yeast and application for inhibitor screening

“…Further metabolism of PE by parasite phospholipases could produce phosphoethanolamine and possibly ethanolamine. Human malaria parasites use a unique phosphoethanolamine methyltransferase PMT for synthesis of phosphocholine from phosphoethanolamine (Pessi et al, 2004;Pessi and Ben Mamoun, 2006;Bobenchik et al, 2010;2011;Bezsonova et al, 2013). The phosphocholine generated from the latter pathway can subsequently be metabolized to CDP-choline and PC.…”

Structure, Function and Inhibition of the Phosphoethanolamine Methyltransferases of the Human Malaria Parasites Plasmodium vivax and Plasmodium knowlesi

“…Structural analysis by NMR and crystallography characterized the nature and specificity of the interactions between PfPMT and substrates/inhibitors 9 24 25 26 27 . Initial residue assignment of PfPMT by NMR made it possible to characterize the interaction between PfPMT and AQ as well as it structural analog chloroquine (CQ) 9 .…”

“…These studies further allowed modeling of AQ on the structure of the enzyme 9 . 1 H, 13 C and 15 N chemical shifts were assigned to elucidate interactions of the enzyme with its substrate and inhibitors to enable determination of a solution structure of PfPMT 24 . Subsequent studies by Lee and colleagues helped solved the structure of the enzyme and provided detailed information about the active site and how the enzyme interacts with its substrates and product 25 .…”

“…Subsequent studies by Lee and colleagues helped solved the structure of the enzyme and provided detailed information about the active site and how the enzyme interacts with its substrates and product 25 . While the structural model proposed by Bobenchik and colleagues predicted a single site of binding of amodiaquine to PfPMT at the entrance of the phosphobase site 9 21 24 , structural analysis by Lee and colleagues suggested that AQ binds at two different allosteric sites 25 . One site is similar to the previously reported by Bobenchik and colleagues at the entrance of the phosphobase site and another site at the surface of the enzyme 25 .…”