Mevalonate 5-diphosphate mediates ATP binding to the mevalonate diphosphate decarboxylase from the bacterial pathogen Enterococcus faecalis
| Autor: | Lake N. Paul, Chun-Liang Chen, James C. Mermoud, Calvin Steussy, Cynthia V. Stauffacher |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Carboxy-Lyases 030106 microbiology Mevalonic Acid Mevalonic acid Biology Crystallography X-Ray Biochemistry Enterococcus faecalis Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Adenosine Triphosphate Hemiterpenes Organophosphorus Compounds Bacterial Proteins Amino Acid Sequence Binding site Molecular Biology chemistry.chemical_classification Binding Sites Isothermal titration calorimetry Cell Biology biology.organism_classification Lyase Kinetics 030104 developmental biology Enzyme chemistry Protein Structure and Folding Mevalonate pathway Adenosine triphosphate |
| Zdroj: | Journal of Biological Chemistry. 292:21340-21351 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m117.802223 |
| Popis: | The mevalonate pathway produces isopentenyl diphosphate (IPP), a building block for polyisoprenoid synthesis, and is a crucial pathway for growth of the human bacterial pathogen Enterococcus faecalis. The final enzyme in this pathway, mevalonate diphosphate decarboxylase (MDD), acts on mevalonate diphosphate (MVAPP) to produce IPP while consuming ATP. This essential enzyme has been suggested as a therapeutic target for the treatment of drug-resistant bacterial infections. Here, we report functional and structural studies on the mevalonate diphosphate decarboxylase from E. faecalis (MDDEF). The MDDEF crystal structure in complex with ATP (MDDEF–ATP) revealed that the phosphate-binding loop (amino acids 97–105) is not involved in ATP binding and that the phosphate tail of ATP in this structure is in an outward-facing position pointing away from the active site. This suggested that binding of MDDEF to MVAPP is necessary to guide ATP into a catalytically favorable position. Enzymology experiments show that the MDDEF performs a sequential ordered bi-substrate reaction with MVAPP as the first substrate, consistent with the isothermal titration calorimetry (ITC) experiments. On the basis of ITC results, we propose that this initial prerequisite binding of MVAPP enhances ATP binding. In summary, our findings reveal a substrate-induced substrate-binding event that occurs during the MDDEF-catalyzed reaction. The disengagement of the phosphate-binding loop concomitant with the alternative ATP-binding configuration may provide the structural basis for antimicrobial design against these pathogenic enterococci. |
| Databáze: | OpenAIRE |
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