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Isolation and immunobiochemical characterization of a major allergen (65 kDa) from Fusarium equiseti
1Centre for Biochemical Technology, Delhi, India.
Abstract:
Fusarium equiseti is one of the most important species in the class Deuteromycetes (Fungi Imperfecti). For proper diagnosis and immunotherapy, isolation and characterization of allergens of F. equiseti are necessary. In the present study, culture filtrate (CF) extract of F. equiseti was resolved into 35-37 bands on isoelectric focusing pI (3-9) and SDS-PAGE (mol. wt. 10-100 kDa). Most of them were glycoproteins, as identified by PAS staining. F. equiseti CF revealed 15 allergenic proteins on immunoblot with an allergic serum pool. It was fractionated into nine fractions (I-IX) on a Superose-12 column by FPLC. Fraction IV (65 kDa) and fraction VI (25 kDa) were found to be highly allergenic by IgE ELISA. A 65-kDa protein was observed as a major allergen because it was recognized by most of the patient sera on immunoblot. After elution from SDS-PAGE gel, it gave two bands of pI 7.4 and 6.0. Inhibition in IgE-binding components of F. equiseti CF with CF extracts of F. solani and F. moniliforme by immunoprint inhibition assay indicated the allergenicity shared between the extracts of Fusarium species. Data suggested that the 65-kDa is the major allergen in the Fusarium species and can be used for the treatment of allergic patients.
Insights
Researchers identified key allergens from Fusarium equiseti, a common fungus. A 65-kDa protein was found to be a major allergen, potentially useful for diagnosing and treating fungal allergies.
Area of Science:
- Mycology
- Immunology
- Allergen research
Background:
- Fusarium equiseti is a significant fungus.
- Identifying its allergens is crucial for diagnosis and immunotherapy.
Purpose of the Study:
- To isolate and characterize allergens from Fusarium equiseti.
- To identify major allergenic components for potential therapeutic use.
Main Methods:
- Culture filtrate (CF) extract analyzed by isoelectric focusing and SDS-PAGE.
- Proteins identified as glycoproteins using PAS staining.
- Allergenic proteins detected by immunoblotting with allergic serum.
- Fractionation using FPLC and IgE ELISA for allergenicity assessment.
Main Results:
- F. equiseti CF contained 35-37 protein bands (10-100 kDa), mostly glycoproteins.
- 15 allergenic proteins were identified.
- A 65-kDa protein in Fraction IV was identified as the major allergen, recognized by most patient sera.
- Cross-reactivity suggested shared allergens among Fusarium species.
Conclusions:
- A 65-kDa protein is the major allergen in Fusarium species.
- This major allergen shows potential for diagnosing and treating allergic reactions to Fusarium.
- Allergenicity is shared across different Fusarium species.