Determination of phosphate-activated glutaminase activity and its kinetics in mouse tissues using metabolic mapping (quantitative enzyme histochemistry).
Bottom Line: PAG activity was decreased to 22% in the presence of 2 mM 6-diazo-5-oxo-L-norleucine.When compared with liver, kidney and brain, other tissues showed 3-fold to 6-fold less PAG activity.In conclusion, PAG is mainly active in mouse kidney, brain and liver, and shows different kinetics depending on which type of PAG is expressed.
Affiliation: Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (DB, WT, CJFVN).Show MeSH
License 1 - License 2 - License 3
Mentions: A method for metabolic mapping of PAG activity is currently not available. Therefore, we developed a novel method with GDH as an auxiliary enzyme using a two-step reaction (Fig. 2; Botman et al. In Press; Van Noorden and Frederiks 1992). Metabolic mapping with a two-step reaction has been used before for other enzymes that also do not use NAD+ or NADP+ as coenzyme to visualize their activity by using the activity of an enzyme that can convert a tetrazolium salt into formazan (Van Noorden and Frederiks 1992).
Affiliation: Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (DB, WT, CJFVN).