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Horse heart acylphosphatase: purification and characterization
The Italian Journal of Biochemistry
|March 1, 1985
Summary
Horse heart acylphosphatase was rapidly purified using affinity chromatography. This enzyme is biochemically identical to acylphosphatase found in horse skeletal muscle, despite lower abundance of certain molecular forms in the heart.
Area of Science:
- Biochemistry
- Enzymology
- Protein Purification
Background:
- Acylphosphatase is an enzyme found in various tissues.
- Understanding tissue-specific enzyme forms is crucial for biochemical research.
- Previous studies focused on acylphosphatase from skeletal muscle.
Purpose of the Study:
- To rapidly purify horse heart acylphosphatase.
- To investigate the molecular forms of horse heart acylphosphatase.
- To compare heart acylphosphatase with skeletal muscle acylphosphatase.
Main Methods:
- Affinity chromatography utilizing anti-horse muscle acylphosphatase antibodies immobilized on Sepharose 4B.
- Purification of acylphosphatase from horse heart tissue.
- Biochemical characterization including amino acid composition, tryptic fingerprinting, molecular weight determination, and kinetic parameter analysis.
Main Results:
- Horse heart acylphosphatase was purified rapidly and efficiently.
- The enzyme exists as a mixed disulfide with glutathione and an S-S dimer, at lower abundance compared to skeletal muscle.
- Purified heart acylphosphatase exhibited identical amino acid composition, tryptic fingerprint, molecular weight, and kinetic parameters to skeletal muscle acylphosphatase.
Conclusions:
- The acylphosphatase present in horse heart is biochemically indistinguishable from that found in horse skeletal muscle.
- The purification method provides an efficient means to isolate acylphosphatase.
- Differences in the abundance of molecular forms suggest potential tissue-specific regulation or post-translational modifications.