Analysis of enzyme activities.
The evaluation of enzyme activities, especially their capacities, represents an important step towards the modelling of biochemical pathways in living organisms. The implementation of microplate technology enables the determination of up to >50 enzymes in relatively large numbers of samples and in various biological materials. Most of these enzymes are involved in central metabolism and several pathways are entirely covered. Direct or indirect assays can be used, as well as highly sensitive assays, depending on the abundance of the enzymes under study. To exemplify such methods, protocols for UDP-glucose pyrophosphorylase (E.C. 2.7.7.9) operating in real time and for pyrophosphate:fructose-6-phosphate 1-phosphotransferase (E.C. 2.7.1.90) are presented.
UMR 1332, INRA, Villenave d'Ornon cedex, France.
Guillaume
G
Ménard
Benoit
B
Biais
Duyên
D
Prodhomme
Patricia
P
Ballias
Yves
Y
Gibon
D011756:Diphosphates / Q000737:chemistry; D057075:Enzyme Assays / Q000592:standards*; D007700:Kinetics; D010732:Phosphofructokinase-1 / Q000737:chemistry / Q000302:isolation & purification; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification; D010944:Plants / Q000201:enzymology; D012015:Reference Standards; D012996:Solutions; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000737:chemistry / Q000302:isolation & purification
D011756:Diphosphates; D010940:Plant Proteins; D012996:Solutions; C107241:diphosphoric acid; D010732:Phosphofructokinase-1; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase