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Protease activity in cockroach and basidiomycete allergen extracts
S Wongtim1, S B Lehrer, J E Salvaggio
1Department of Medicine, Tulane University School of Medicine, New Orleans, Louisiana.
Allergy Proceedings : the Official Journal of Regional and State Allergy Societies
|July 1, 1993
Summary
Cockroach and mushroom extracts contain proteolytic enzymes that can degrade allergens. Minimizing protease activity during allergen extract preparation is crucial for preserving allergenicity and ensuring accurate diagnostic testing.
Area of Science:
- Biochemistry
- Enzymology
- Allergenology
Background:
- Proteolytic enzymes are present in various biological extracts.
- These enzymes can potentially degrade allergenic proteins, impacting diagnostic accuracy.
- Understanding protease activity in allergen sources is vital for standardization.
Purpose of the Study:
- To quantify the inherent proteolytic activity in cockroach and basidiomycete extracts.
- To investigate the impact of protease inhibitors on this activity.
- To correlate protease activity with allergenicity loss.
Main Methods:
- Assessing proteolytic activity by measuring acid-soluble peptides released from bovine serum albumin using the Lowry method.
- Utilizing reference proteases (Proteinase K, Trypsin, Pronase) for comparison.
- Employing protease inhibitors phenylmethylsulfonyl fluoride and ethylene diaminetetraacetic acid to assess inhibition.
Main Results:
- American cockroach whole-body extract exhibited significant proteolytic activity (330 µg/mL, 13 TEU/mg).
- Extracts from Pleurotus ostreatus spores (230 µg/mL, 0.9 TEU/mg) and caps (112 µg/mL, 0.5 TEU/mg) showed moderate activity.
- Phenylmethylsulfonyl fluoride inhibited cockroach extract activity by 80%, while ethylene diaminetetraacetic acid inhibited Pleurotus spore activity by 95%.
Conclusions:
- Cockroach and certain mushroom extracts possess significant proteolytic activity.
- Protease activity directly correlates with the loss of allergen activity.
- Conditions minimizing proteolysis are recommended for allergen extract preparation and handling to maintain diagnostic reliability.