Projects per year
Abstract
Previous metabolic studies have demonstrated that leishmania parasites are able to synthesise proline from glutamic acid and threonine from aspartic acid. The first committed step in both biosynthetic pathways involves an amino acid kinase, either a glutamate 5‐kinase (G5K; EC2.7.2.11) or an aspartokinase (EC2.7.2.4). Bioinformatic analysis of multiple leishmania genomes identifies a single amino acid‐kinase gene (LdBPK 262740.1) variously annotated as either a putative glutamate or aspartate kinase. To establish the catalytic function of this Leishmania donovani gene product, we have determined the physical and kinetic properties of the recombinant enzyme purified from Escherichia coli. The findings indicate that the enzyme is a bona fide G5K with no activity as an aspartokinase. Tetrameric G5K displays kinetic behaviour similar to its bacterial orthologues and is allosterically regulated by proline, the end product of the pathway. The structure‐activity relationships of proline analogues as inhibitors are broadly similar to the bacterial enzyme. However, unlike G5K from E. coli, leishmania G5K lacks a C‐terminal PUA (pseudouridine synthase and archaeosine transglycosylase) domain and does not undergo higher oligomerisation in the presence of proline. Gene replacement studies are suggestive, but not conclusive that G5K is essential.
Original language | English |
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Pages (from-to) | 2662-2678 |
Number of pages | 17 |
Journal | FEBS Journal |
Volume | 285 |
Issue number | 14 |
Early online date | 19 May 2018 |
DOIs | |
Publication status | Published - Jul 2018 |
Keywords
- Leishmania
- glutamate 5-kinase
- RCWT
- wild/type leishmania overexpressing G5K
- SDR
- single drug resistant line
- DDR
- double drug resistant line
- PUA domain
- pseudouridine synthase and archaeosine transglycosylase domain
- P5CS
- UTR
- untranslated region
- inhibitors
- biochemical pathway
- proline biosynthesis
- drug target
ASJC Scopus subject areas
- Molecular Biology
- Biochemistry
- Cell Biology
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- 1 Finished
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Aref#d: 18185. Characterization and validation of drug targets in the Kinetoplastida (Principal Research Fellowship/Programme Grant)
Fairlamb, A. (Investigator)
1/10/06 → 30/09/17
Project: Research
Profiles
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Fairlamb, Alan
- Biological Chemistry and Drug Discovery - Associate Staff of Biochemistry (Consulting)
Person: Associate Staff