190079-18-6Relevant articles and documents
Observation of thiamin-bound intermediates and microscopic rate constants for their interconversion on 1-deoxy- d -xylulose 5-phosphate synthase: 600-Fold rate acceleration of pyruvate decarboxylation by d -glyceraldehyde-3-phosphate.
Patel, Hetalben,Nemeria, Natalia S.,Jordan, Frank,Brammer, Leighanne A.,Freel Meyers, Caren L.
, p. 18374 - 18379,6 (2012)
The thiamin diphosphate (ThDP)-dependent enzyme 1-deoxy-d-xylulose 5-phosphate (DXP) synthase carries out the condensation of pyruvate as a 2-hydroxyethyl donor with d-glyceraldehyde-3-phosphate (d-GAP) as acceptor forming DXP. Toward understanding catalysis of this potential anti-infective drug target, we examined the pathway of the enzyme using steady state and presteady state kinetic methods. It was found that DXP synthase stabilizes the ThDP-bound predecarboxylation intermediate formed between ThDP and pyruvate (C2α-lactylThDP or LThDP) in the absence of d-GAP, while addition of d-GAP enhanced the rate of decarboxylation by at least 600-fold. We postulate that decarboxylation requires formation of a ternary complex with both LThDP and d-GAP bound, and the central enzyme-bound enamine reacts with d-GAP to form DXP. This appears to be the first study of a ThDP enzyme where the individual rate constants could be evaluated by time-resolved circular dichroism spectroscopy, and the results could have relevance to other ThDP enzymes in which decarboxylation is coupled to a ligation reaction. The acceleration of the rate of decarboxylation of enzyme-bound LThDP in the presence of d-GAP suggests a new approach to inhibitor design.
An improved preparation of D-glyceraldehyde 3-phosphate and its use in the synthesis of 1-deoxy-D-xylulose 5-phosphate
Li, Heng,Tian, Jie,Wang, Hui,Yang, Shao-Qing,Gao, Wen-Yun
, p. 1745 - 1750 (2010)
D-Glyceraldehyde 3-phosphate (=D-GAP; 2) was prepared by an improved chemical method (Scheme 2), and it was then employed to synthesize 1-deoxy-d-xylulose 5-phosphate (=DXP; 3) which is enzymatically one of the key intermediates in the MEP (4) terpenoid b
Formation of 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol from 2-C- methyl-D-erythritol 4-phosphate by 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase, a new enzyme in the nonmevalonate pathway
Kuzuyama, Tomohisa,Takagi, Motoki,Kaneda, Kazuhide,Dairi, Tohru,Seto, Haruo
, p. 703 - 706 (2000)
2-C-Methyl,D-erythritol 4-phosphate is transformed to 4-(cytidine 5'- diphospho)-2-C-methyl-D-erythritol in the presence of cytidine 5'- triphosphate by a novel Escherichia coli enzyme, 2-C-methyl-D-erythritol 4- phosphate cytidylyltransferase, involved in the nonmevalonate pathway. (C) 2000 Elsevier Science Ltd.
DXP synthase-catalyzed c-n bond formation: Nitroso substrate specificity studies guide selective inhibitor design
Morris, Francine,Vierling, Ryan,Boucher, Lauren,Bosch, Juergen,Freel Meyers, Caren L.
, p. 1309 - 1315 (2013/08/23)
1-Deoxy-D-xylulose 5-phosphate (DXP) synthase catalyzes the first step in the nonmammalian isoprenoid biosynthetic pathway to form DXP from pyruvate and D-glyceraldehyde 3-phosphate (D-GAP) in a thiamin diphosphate-dependent manner. Its unique structure and mechanism distinguish DXP synthase from its homologues and suggest that it should be pursued as an anti-infective drug target. However, few reports describe any development of selective inhibitors of this enzyme. Here, we reveal that DXP synthase catalyzes C-N bond formation and exploit aromatic nitroso substrates as active site probes. Substrate specificity studies reveal a high affinity of DXP synthase for aromatic nitroso substrates compared to the related ThDP-dependent enzyme pyruvate dehydrogenase (PDH). Results from inhibition and mutagenesis studies indicate that nitroso substrates bind to E. coli DXP synthase in a manner distinct from that of D-GAP. Our results suggest that the incorporation of aryl acceptor substrate mimics into unnatural bisubstrate analogues will impart selectivity to DXP synthase inhibitors. As a proof of concept, we show selective inhibition of DXP synthase by benzylacetylphosphonate (BnAP).