Protease activity in cockroach and basidiomycete allergen extracts
| Autor: | John E. Salvaggio, Samuel B. Lehrer, Horner We, Wongtim S |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Pulmonary and Respiratory Medicine
Psilocybe cubensis medicine.medical_treatment Cockroaches Pronase Puffball Polyporaceae Radioallergosorbent Test Immunology and Allergy Medicine Animals Humans Protease Inhibitors Trypsin Calvatia cyathiformis Pleurotus Protease Chromatography biology business.industry Basidiomycota fungi Serine Endopeptidases General Medicine Allergens biology.organism_classification Pleurotus ostreatus Endopeptidase K business Agaricales medicine.drug |
| Zdroj: | Allergy proceedings : the official journal of regional and state allergy societies. 14(4) |
| ISSN: | 1046-9354 |
| Popis: | Inherent proteolytic activity was estimated in cockroach and basidiomycete extracts by quantifying acid soluble peptides that were released by incubating extracts with 1% bovine serum albumin as measured by Lowry (Sigma). Reference proteases released 740 (Proteinase K, 0.1 U), 248 (Trypsin, 1.0 U), and 533 micrograms/ml (Pronase, 0.5 U) of soluble peptides. American whole body cockroach extract (0.1 mg dry weight) released 330 micrograms/ml of soluble peptides, representing 13 trypsin equivalent units (TEU)/mg. Extracts from spores of the mushroom Pleurotus ostreatus released 230 micrograms/ml (0.9 TEU/mg) and Pleurotus cap extract released 112 micrograms/ml (0.5 TEU/mg). Mycelium of Pleurotus and the mushroom Psilocybe cubensis and spores of Psilocybe and the puffball Calvatia cyathiformis showed negligible amounts of proteolytic activity. The protease inhibitor phenylmethylsulfonyl flouride reduced the proteolytic activity of American whole body cockroach extract by 80% (@1 mM) and the inhibitor ethylene diaminetetraacetic acid inhibited the proteolytic activity of Pleurotus spores by 95% (@1 mM). Loss of allergen activity as determined by RAST inhibition and immunoprinting correlated with protease activity. Thus, in the preparation and handling of allergen extracts, one should employ conditions that minimize proteolysis. |
| Databáze: | OpenAIRE |
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