title: Measurement of carbon flux through the MEP pathway for isoprenoid synthesis by (31)P-NMR spectroscopy after specific inhibition of 2-C-methyl-d-erythritol 2,4-cyclodiphosphate reductase. Effect of light and temperature.
abstract: The methylerythritol 4-phosphate (MEP) and the mevalonate pathways are the unique synthesis routes for the precursors of all isoprenoids. An original mean to measure the carbon flux through the MEP pathway in plants is proposed by using cadmium as a total short-term inhibitor of 2-C-methyl-d-erythritol 2,4-cyclodiphosphate (MEcDP) reductase (GcpE) and measuring the accumulation rate of its substrate MEcDP by (31) P-NMR spectroscopy. The MEP pathway metabolic flux was determined in spinach (Spinacia oleracea), pea (Pisum sativum), Oregon grape (Mahonia aquifolium) and boxwood (Buxus sempervirens) leaves. In spinach, flux values were compared with the synthesis rate of major isoprenoids. The flux increases with light intensity (fourfold in the 200-1200 µmol m(-2) s(-1) PPFR range) and temperature (sevenfold in the 25-37 °C range). The relationship with the light and the temperature dependency of isoprenoid production downstream of the MEP pathway is discussed.
authors: CEA, IRTSV, Laboratoire de Physiologie Cellulaire Végétale, UMR 5168, F-38054 Grenoble, France.
authors: Gaëlle
authors: G
authors: Mongélard
authors: Myriam
authors: M
authors: Seemann
authors: Anne-Marie
authors: AM
authors: Boisson
authors: Michel
authors: M
authors: Rohmer
authors: Richard
authors: R
authors: Bligny
authors: Corinne
authors: C
authors: Rivasseau
mesh_terms: D031235:Buxus / Q000378:metabolism; D002104:Cadmium / Q000494:pharmacology; D057486:Carbon Cycle; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004896:Erythritol / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D008027:Light; D009682:Magnetic Resonance Spectroscopy; D028143:Mahonia / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D008798:Mevalonic Acid / Q000378:metabolism; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism; D018532:Pisum sativum / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D010944:Plants / Q000378:metabolism*; D018724:Spinacia oleracea / Q000378:metabolism; D013696:Temperature; D013729:Terpenes / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D010761:Phosphorus Radioisotopes; D013729:Terpenes; C443879:methyl-D-erythritol 2,4-cyclodiphosphate; D002104:Cadmium; D010088:Oxidoreductases; D004896:Erythritol; D008798:Mevalonic Acid