An assay for arginase in chicken kidney
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Published on: November 5, 2014
1Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853.
Abstract:
1. The enzyme arginase in chicken kidney is associated with mitochondria and the mitochondrial membranes must be disrupted to obtain maximum activity. 2. When the membranes were disrupted by sonication, approximately 30% higher 2. When the membranes were disrupted by sonication, approximately 30% higher arginase activity was observed than with the nonsonicated samples. 3. The optimum pH for assay of chick kidney arginase was 9.7-9.8. Prior heat treatment with Mn2+ decreased arginase activity. 4. Highest enzyme activity was obtained by using sonicated preparations and measuring initial reaction velocity during the first 1-2 min of incubation.
Insights
Maximizing chicken kidney arginase activity requires disrupting mitochondrial membranes, with sonication yielding a 30% increase. Optimal assay conditions involve a pH of 9.7-9.8 and measuring initial reaction velocity.
Area of Science:
- Biochemistry
- Enzymology
- Cellular Biology
Background:
- Arginase is a key enzyme in the urea cycle.
- Its localization within chicken kidney mitochondria influences activity.
- Understanding enzyme kinetics is crucial for biochemical studies.
Purpose of the Study:
- To determine the optimal conditions for assaying chicken kidney arginase activity.
- To investigate the effect of mitochondrial membrane disruption on enzyme activity.
- To establish reliable methods for measuring arginase kinetics.
Main Methods:
- Chicken kidney mitochondria were isolated.
- Mitochondrial membranes were disrupted using sonication.
- Arginase activity was assayed at various pH levels.
- Initial reaction velocities were measured under optimal conditions.
Main Results:
- Sonication of mitochondrial membranes increased arginase activity by approximately 30%.
- The optimal pH for chicken kidney arginase assay was determined to be between 9.7 and 9.8.
- Prior heat treatment with Mn2+ resulted in decreased arginase activity.
- Highest enzyme activity was achieved with sonicated preparations measuring initial reaction velocity within 1-2 minutes.
Conclusions:
- Disruption of mitochondrial membranes is essential for maximal arginase activity in chicken kidneys.
- Sonication is an effective method for membrane disruption, significantly enhancing enzyme yield.
- Precise control of pH and reaction time is critical for accurate kinetic analysis of chicken kidney arginase.
