Biochemical and Biophysical Analysis of Phosphoethanolamine N-methyltransferase in the Protozoan Parasites
Disciplines
Amino Acids, Peptides, and Proteins | Biochemistry | Medicinal and Pharmaceutical Chemistry | Molecular Biology | Other Biochemistry, Biophysics, and Structural Biology | Pharmaceutical Preparations | Structural Biology
Abstract (300 words maximum)
Phosphatidylcholine (PtdCho) is one of the essential lipids present in the cell membranes of plants, animals, and prokaryotes. The biosynthesis of phosphatidylcholine involves the methylation of Phosphoethanolamine (pEA) to phosphocholine (pCho). This methylation process is mediated by the enzyme called Phosphoethanolamine N-methyltransferase (PMT). PMT is available in plants, nematodes, and some protozoan parasites, including Plasmodium and Eimeria species. Among them, Eimeria parasites are abundant in the world and responsible for parasitic infection. Eimeria parasites cause coccidiosis in chickens and other birds, leading to severe economic loss for the poultry industry. The treatment is expensive, and the price of poultry-based protein is getting high and ultimately humans are suffering. Only a few drugs are available to treat this disease, but resistance and allergic reactions are reported. Thus, the identification of biochemical targets and the development of antiparasitic drugs have great importance. PMT plays a vital role in their cell growth and development. In this study, a series of biochemical and biophysical analyses, including enzyme kinetics, protein-ligand binding, and X-ray crystallography, provides a foundation for drug development. Specifically, we determined the three-dimensional structures of EtPMT at a 2.9 Å resolution. Additionally, functional characterization was performed using enzyme and binding assays, which revealed valuable properties of EtPMT, such as its Km and Kd values. The QuikChange site-directed mutagenesis kit was employed to gain detailed insights into the reaction mechanism of PMT. The next step involves fragment-based screening and molecular docking, which will aid in developing novel compounds targeting PMT as potential antiparasitic drugs.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Dr. Soon Goo Lee
Biochemical and Biophysical Analysis of Phosphoethanolamine N-methyltransferase in the Protozoan Parasites
Phosphatidylcholine (PtdCho) is one of the essential lipids present in the cell membranes of plants, animals, and prokaryotes. The biosynthesis of phosphatidylcholine involves the methylation of Phosphoethanolamine (pEA) to phosphocholine (pCho). This methylation process is mediated by the enzyme called Phosphoethanolamine N-methyltransferase (PMT). PMT is available in plants, nematodes, and some protozoan parasites, including Plasmodium and Eimeria species. Among them, Eimeria parasites are abundant in the world and responsible for parasitic infection. Eimeria parasites cause coccidiosis in chickens and other birds, leading to severe economic loss for the poultry industry. The treatment is expensive, and the price of poultry-based protein is getting high and ultimately humans are suffering. Only a few drugs are available to treat this disease, but resistance and allergic reactions are reported. Thus, the identification of biochemical targets and the development of antiparasitic drugs have great importance. PMT plays a vital role in their cell growth and development. In this study, a series of biochemical and biophysical analyses, including enzyme kinetics, protein-ligand binding, and X-ray crystallography, provides a foundation for drug development. Specifically, we determined the three-dimensional structures of EtPMT at a 2.9 Å resolution. Additionally, functional characterization was performed using enzyme and binding assays, which revealed valuable properties of EtPMT, such as its Km and Kd values. The QuikChange site-directed mutagenesis kit was employed to gain detailed insights into the reaction mechanism of PMT. The next step involves fragment-based screening and molecular docking, which will aid in developing novel compounds targeting PMT as potential antiparasitic drugs.