Measuring the activity of 1-deoxy-D-xylulose 5-phosphate synthase, the first enzyme in the MEP pathway, in plant extracts.
The first enzyme in the methylerythritol phosphate (MEP) pathway is 1-deoxy-D-xylulose 5-phosphate (DXP) synthase (DXS). As such this enzyme is considered to be important in the control of plastidial isoprenoid production. Measuring the activity of DXS in plant extracts is therefore crucial to understanding the regulation of the MEP pathway. Due to the relatively low amounts of DXS, the activity of this enzyme can only be measured using highly sensitive analytical equipment. Here, a method is described to determine the DXS enzyme activity in a crude plant extract, by measuring DXP production directly using high performance liquid chromatography linked to a tandem triple quadrupole mass spectrometry detector (LC-MS/MS).
Department of Biochemistry, Max Planck Institute for Chemical Ecology, Hans-Knöll-Str. 8, 07745, Jena, Germany, lwright@ice.mpg.de.
Louwrance P
LP
Wright
Michael A
MA
Phillips
D017360:Arabidopsis / Q000201:enzymology* / Q000378:metabolism; D002853:Chromatography, Liquid; D057075:Enzyme Assays / Q000379:methods*; D004896:Erythritol / Q000378:metabolism*; D010936:Plant Extracts / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D014166:Transferases / Q000302:isolation & purification / Q000378:metabolism*
D010936:Plant Extracts; D014166:Transferases; C109461:deoxyxylulose-5-phosphate synthase; D004896:Erythritol