Mechanistic binding insights for 1-deoxy-d-Xylulose-5-Phosphate synthase, the enzyme catalyzing the first reaction of isoprenoid biosynthesis in the malaria-causing protists, Plasmodium falciparum and Plasmodium vivax
| Autor: | David J. Merkler, Matthew R. Battistini, Christopher Shoji, Leonid Breydo, Sumit Handa |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Molecular Sequence Data Plasmodium falciparum Plasmodium vivax Protozoan Proteins lac operon Sequence alignment Biology Glyceraldehyde 3-Phosphate Plasmodium Article 03 medical and health sciences chemistry.chemical_compound Transferases Catalytic Domain Pyruvic Acid parasitic diseases Amino Acid Sequence Cloning Molecular chemistry.chemical_classification 030102 biochemistry & molecular biology ATP synthase biology.organism_classification Recombinant Proteins Kinetics 030104 developmental biology Enzyme Biochemistry chemistry biology.protein Glyceraldehyde 3-phosphate Sequence Alignment Biotechnology |
| Zdroj: | Protein Expression and Purification. 120:16-27 |
| ISSN: | 1046-5928 |
| Popis: | We have successfully truncated and recombinantly-expressed 1-deoxy-D-xylulose-5-phosphate synthase (DXS) from both Plasmodium vivax and Plasmodium falciparum. We elucidated the order of substrate binding for both of these ThDP-dependent enzymes using steady-state kinetic analyses, dead-end inhibition, and intrinsic tryptophan fluorescence titrations. Both enzymes adhere to a random sequential mechanism with respect to binding of both substrates: pyruvate and D-glyceraldehyde-3-phosphate. These findings are in contrast to other ThDP-dependent enzymes, which exhibit classical ordered and/or ping-pong kinetic mechanisms. A better understanding of the kinetic mechanism for these two Plasmodial enzymes could aid in the development of novel DXS-specific inhibitors that might prove useful in treatment of malaria. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect