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Pulmonary angiotensin-converting enzyme. Structural and catalytic properties
This study details the purification and characterization of rabbit lung angiotensin-converting enzyme (ACE), a glycoprotein containing zinc. The purified enzyme efficiently hydrolyzes angiotensin I and bradykinin, with activity modulated by chloride ions and specific peptides.
Area of Science:
- Biochemistry
- Enzymology
- Proteomics
Background:
- Angiotensin-converting enzyme (ACE) plays a crucial role in the renin-angiotensin system.
- Understanding ACE structure and function is vital for cardiovascular research.
Purpose of the Study:
- To purify and characterize angiotensin-converting enzyme from rabbit lung.
- To investigate the enzyme's molecular properties, substrate specificity, and kinetic parameters.
Main Methods:
- Enzyme purification using DEAE-cellulose, calcium phosphate gel, Sephadex G-200, and lectin affinity chromatography.
- Molecular weight determination via equilibrium sedimentation and SDS-PAGE.
- Amino acid and carbohydrate composition analysis.
- Kinetic studies using hippurylhistidylleucine, angiotensin I, and bradykinin as substrates.
Main Results:
- Purified ACE to apparent homogeneity with an 11% yield.
- Determined molecular weight of ~129,000 Da, with glycoprotein nature contributing to higher estimates.
- Identified zinc as a component and characterized kinetic parameters (Km, turnover numbers) for substrates.
- Demonstrated inhibition by EDTA and bradykinin-potentiating peptides, and competitive inhibition by other peptides.
Conclusions:
- Rabbit lung ACE is a zinc-containing glycoprotein with significant enzymatic activity.
- The enzyme's activity is dependent on chloride ions and sensitive to specific peptide inhibitors.
- Characterization provides insights into ACE's biochemical properties and potential regulatory mechanisms.
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