Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 1866-1874 |
| Seitenumfang | 9 |
| Fachzeitschrift | CHEMBIOCHEM |
| Jahrgang | 6 |
| Ausgabenummer | 10 |
| Publikationsstatus | Veröffentlicht - 1 Okt. 2005 |
| Extern publiziert | Ja |
Abstract
The inhibition of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) by fosmidomycin was studied by using a kinetic assay based on the consumption of NADPH and synthetic substrate. Fosmidomycin is a slow tight-binding inhibitor of DXR that shows strong negative cooperativity (|h| = 0.3) in binding. Cooperativity is displayed during the initial (weak, K0.5 = 10 μM) binding event and does not change as the binding tightens to the equilibrium value of 0.9 nM over a period of seconds to minutes. A series of fosmidomycin fragments was examined, but all showed much weaker inhibition, in the mM range. A series of cyclic fosmidomycin analogues was also synthesised and tested, but these showed high-μM binding at best. None of the synthetic compounds showed time-dependent inhibition. We concluded that the slow tight-binding behaviour, and perhaps also cooperativity, are mediated by significant reorganisation of the active site upon fosmidomycin binding. This makes the rational design of new inhibitors of DXR difficult at best.
ASJC Scopus Sachgebiete
- Biochemie, Genetik und Molekularbiologie (insg.)
- Biochemie
- Biochemie, Genetik und Molekularbiologie (insg.)
- Molekularmedizin
- Biochemie, Genetik und Molekularbiologie (insg.)
- Molekularbiologie
- Chemie (insg.)
- Organische Chemie
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in: CHEMBIOCHEM, Jahrgang 6, Nr. 10, 01.10.2005, S. 1866-1874.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - A fragment-based approach to understanding inhibition of 1-deoxy-D-xylulose-5-phosphate reductoisomerase
AU - Mercklé, Ludovic
AU - De Andrés-Gómez, Ana
AU - Dick, Bethany
AU - Cox, Russell J.
AU - Godfrey, Christopher R.A.
PY - 2005/10/1
Y1 - 2005/10/1
N2 - The inhibition of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) by fosmidomycin was studied by using a kinetic assay based on the consumption of NADPH and synthetic substrate. Fosmidomycin is a slow tight-binding inhibitor of DXR that shows strong negative cooperativity (|h| = 0.3) in binding. Cooperativity is displayed during the initial (weak, K0.5 = 10 μM) binding event and does not change as the binding tightens to the equilibrium value of 0.9 nM over a period of seconds to minutes. A series of fosmidomycin fragments was examined, but all showed much weaker inhibition, in the mM range. A series of cyclic fosmidomycin analogues was also synthesised and tested, but these showed high-μM binding at best. None of the synthetic compounds showed time-dependent inhibition. We concluded that the slow tight-binding behaviour, and perhaps also cooperativity, are mediated by significant reorganisation of the active site upon fosmidomycin binding. This makes the rational design of new inhibitors of DXR difficult at best.
AB - The inhibition of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) by fosmidomycin was studied by using a kinetic assay based on the consumption of NADPH and synthetic substrate. Fosmidomycin is a slow tight-binding inhibitor of DXR that shows strong negative cooperativity (|h| = 0.3) in binding. Cooperativity is displayed during the initial (weak, K0.5 = 10 μM) binding event and does not change as the binding tightens to the equilibrium value of 0.9 nM over a period of seconds to minutes. A series of fosmidomycin fragments was examined, but all showed much weaker inhibition, in the mM range. A series of cyclic fosmidomycin analogues was also synthesised and tested, but these showed high-μM binding at best. None of the synthetic compounds showed time-dependent inhibition. We concluded that the slow tight-binding behaviour, and perhaps also cooperativity, are mediated by significant reorganisation of the active site upon fosmidomycin binding. This makes the rational design of new inhibitors of DXR difficult at best.
KW - Antibiotics
KW - Enzymes
KW - Fosmidomycin
KW - Inhibitors
KW - Simulated docking
UR - http://www.scopus.com/inward/record.url?scp=26944489968&partnerID=8YFLogxK
U2 - 10.1002/cbic.200500061
DO - 10.1002/cbic.200500061
M3 - Article
C2 - 16116659
AN - SCOPUS:26944489968
VL - 6
SP - 1866
EP - 1874
JO - CHEMBIOCHEM
JF - CHEMBIOCHEM
SN - 1439-4227
IS - 10
ER -