Related Experiment Videos
Human skin tryptase: purification, partial characterization and comparison with human lung tryptase.
I T Harvima1, N M Schechter, R J Harvima
1Department of Dermatology, University of Kuopio, Finland.
Biochimica Et Biophysica Acta
|November 2, 1988
Summary
Human skin and lung tryptases are closely related trypsin-like proteinases. Purification revealed identical molecular sizes and properties, suggesting mast cells contain similar enzymes.
Area of Science:
- Biochemistry
- Enzymology
- Cell Biology
Background:
- Human skin tryptase is a serine protease found in mast cells.
- Tryptases play roles in various physiological and pathological processes.
Purpose of the Study:
- To purify and characterize human skin tryptase.
- To compare human skin tryptase with human lung tryptase.
Main Methods:
- Stepwise salt extraction, hydrophobic affinity chromatography, gel filtration, and ion exchange chromatography were used for purification.
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration were used to determine molecular weight.
- Enzyme kinetics and immunodiffusion were employed for characterization and comparison.
Main Results:
- Human skin tryptase was purified 448-fold.
- The native enzyme (Mr 120,000) consists of subunits (Mr 34,000 and 38,000).
- Purified skin and lung tryptases exhibited identical molecular sizes, kinetic parameters, inhibition profiles, and immunological cross-reactivity.
Conclusions:
- Human skin and lung tryptases are closely related, potentially identical trypsin-like proteinases.
- Mast cells in both skin and lung likely contain similar enzymes.
- These findings contribute to understanding tryptase function and mast cell heterogeneity.