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 ascertain whether the auxiliary GDH activity of the GDH films between cryostat sections and glass slides were available throughout the tissue sections in a zero order manner (i.e., not rate limiting), we determined the distribution of GDH activity over the glass slides (Fig. 3A). GDH activity on the glass slides showed little variation over the entire set of films, indicating that GDH was equally distributed across the glass slides. Furthermore, no significant differences in absorbance (i.e., GDH activity) was found between the tested films (one-way ANOVA at α=0.01).
Affiliation: Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (DB, WT, CJFVN).