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Solubilization of particle-linked urate oxidase by different agents
Acta Physiologica Latino Americana
|January 1, 1975
Summary
Researchers explored methods to extract urate oxidase (an enzyme) from cell fractions. High alkaline pH, freezing, and calcium chloride effectively solubilized the enzyme, suggesting electrostatic forces maintain its structure.
Area of Science:
- Biochemistry
- Cell Biology
- Enzymology
Background:
- Urate oxidase is an enzyme often found in subcellular fractions.
- Understanding enzyme extraction is crucial for biochemical studies.
- Previous research observed crystalloid structures related to urate oxidase.
Purpose of the Study:
- To investigate effective methods for solubilizing and extracting sedimentable urate oxidase.
- To determine agents that can release urate oxidase activity from subcellular fractions.
- To explore the nature of forces responsible for the enzyme's structural integrity.
Main Methods:
- Subcellular fractionation of homogenates to isolate urate oxidase.
- Testing various agents (pH, detergents, ionic strength) for enzyme solubilization.
- Centrifugation to separate solubilized enzyme in supernatants from insoluble pellets.
- Assessing enzyme activity and protein extraction efficiency.
Main Results:
- Urate oxidase was primarily recovered in a high-speed centrifugation pellet.
- Agents like high alkaline pH, Hyamine 2389, and calcium chloride effectively solubilized the enzyme.
- Triton X-100 extracted proteins but left most urate oxidase activity insoluble.
- Extraction efficiencies approached 100% with specific agents, suggesting disruption of structural integrity.
Conclusions:
- Electrostatic forces are likely responsible for maintaining the integrity of urate oxidase-related crystalloids.
- Specific chemical and physical treatments can efficiently release and solubilize urate oxidase.
- The findings provide insights into the biochemical properties and extraction of urate oxidase.