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Determination of phosphate-activated glutaminase activity and its kinetics in mouse tissues using metabolic mapping (quantitative enzyme histochemistry).

Botman D, Tigchelaar W, Van Noorden CJ - J. Histochem. Cytochem. (2014)

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.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (DB, WT, CJFVN).

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Phosphate-activated glutaminase (PAG) activity in brain tissue sections of a mouse in the presence of 0 mM, 0.5 mM, and 2 mM 6-diazo-5-oxo-L-norleucine (DON; PAG-specific inhibitor). PAG activity was inhibited dose-dependently by DON. All test reactions were corrected for nonspecific background staining in the absence of substrate. Error bars indicate SEM (n=10).
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fig5-0022155414551177: Phosphate-activated glutaminase (PAG) activity in brain tissue sections of a mouse in the presence of 0 mM, 0.5 mM, and 2 mM 6-diazo-5-oxo-L-norleucine (DON; PAG-specific inhibitor). PAG activity was inhibited dose-dependently by DON. All test reactions were corrected for nonspecific background staining in the absence of substrate. Error bars indicate SEM (n=10).

Mentions: To confirm the specificity of the PAG activity staining method, PAG activity in brain tissue was examined in the presence of the PAG inhibitor DON (Fig. 5). PAG activity staining was significantly reduced to 22% in the presence of 2 mM DON as compared with the staining in the absence of DON (Students t-test, p<0.0001).


Determination of phosphate-activated glutaminase activity and its kinetics in mouse tissues using metabolic mapping (quantitative enzyme histochemistry).

Botman D, Tigchelaar W, Van Noorden CJ - J. Histochem. Cytochem. (2014)

Phosphate-activated glutaminase (PAG) activity in brain tissue sections of a mouse in the presence of 0 mM, 0.5 mM, and 2 mM 6-diazo-5-oxo-L-norleucine (DON; PAG-specific inhibitor). PAG activity was inhibited dose-dependently by DON. All test reactions were corrected for nonspecific background staining in the absence of substrate. Error bars indicate SEM (n=10).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2 - License 3
Show All Figures
getmorefigures.php?uid=PMC4230542&req=5

fig5-0022155414551177: Phosphate-activated glutaminase (PAG) activity in brain tissue sections of a mouse in the presence of 0 mM, 0.5 mM, and 2 mM 6-diazo-5-oxo-L-norleucine (DON; PAG-specific inhibitor). PAG activity was inhibited dose-dependently by DON. All test reactions were corrected for nonspecific background staining in the absence of substrate. Error bars indicate SEM (n=10).
Mentions: To confirm the specificity of the PAG activity staining method, PAG activity in brain tissue was examined in the presence of the PAG inhibitor DON (Fig. 5). PAG activity staining was significantly reduced to 22% in the presence of 2 mM DON as compared with the staining in the absence of DON (Students t-test, p<0.0001).

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.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (DB, WT, CJFVN).

Show MeSH
Related in: MedlinePlus