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Analysis of enzyme activities.

Authors
  • Ménard, Guillaume
  • Biais, Benoit
  • Prodhomme, Duyên
  • Ballias, Patricia
  • Gibon, Yves
Type
Published Article
Journal
Methods in Molecular Biology
Publication Date
2014
Volume
1090
Pages
249–259
Identifiers
DOI: 10.1007/978-1-62703-688-7_15
PMID: 24222420
Source
Medline
License
Unknown

Abstract

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.

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