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
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Mentions: To determine the reproducibility of the PAG activity staining method and variation within the samples, tissue sections were stained for PAG activity under identical conditions with varied glutamine concentrations (Fig. 6). The staining method showed the highest variation at low glutamine concentrations. Variations in our method of determining PAG activity was not significantly different from other methods, such as the metabolic mapping method to demonstrate GDH activity (analyzed for liver tissue; F-test at α=0.05; data not shown).
Affiliation: Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (DB, WT, CJFVN).