Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10533267 | Analytical Biochemistry | 2005 | 7 Pages |
Abstract
A new method for the determination of the activity of 4-diphosphocytidyl-2-C-methyl-d-erythritol 4-phosphate synthase, the enzyme catalyzing the third reaction of the 2-C-methyl-d-erythritol 4-phosphate pathway for biosynthesis of isoprenoids, is described. This is an end-point assay based on the transformation of inorganic pyrophosphate, one of the products of the reaction, to phosphate by using inorganic pyrophosphatase as auxiliary enzyme. The phosphate formed is reacted then with the dye malachite green to yield a colored product which can be determined spectrophotometrically. The method is easy to perform, sensitive, and robust and can be used in automated high-throughput screening analyses for the search of inhibitors of the enzyme.
Keywords
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Cristobal Bernal, Celia Palacin, Albert Boronat, Santiago Imperial,