Determination of 3-hydroxy-3-methylglutaryl CoA reductase activity in plants.
The enzyme 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase catalyzes the NADPH-mediated reductive deacylation of HMG-CoA to mevalonic acid, which is the first committed step of the mevalonate pathway for isoprenoid biosynthesis. In agreement with its key regulatory role in the pathway, plant HMG-CoA reductase is modulated by many diverse external stimuli and endogenous factors and can be detected to variable levels in every plant tissue. A fine determination of HMG-CoA reductase activity levels is required to understand its contribution to plant development and adaptation to changing environmental conditions. Here, we report a procedure to reliably determine HMG-CoA reductase activity in plants. The method includes the sample collection and homogenization strategies as well as the specific activity determination based on a classical radiochemical assay.
Center for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Campus UAB Bellaterra, Barcelona, Spain, ncampos@ub.edu.
Narciso
N
Campos
Montserrat
M
Arró
Albert
A
Ferrer
Albert
A
Boronat
D017360:Arabidopsis / Q000201:enzymology*; D002855:Chromatography, Thin Layer; D057075:Enzyme Assays / Q000379:methods*; D006903:Hydroxymethylglutaryl CoA Reductases / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity
D006903:Hydroxymethylglutaryl CoA Reductases