Production and characterization of a noncytotoxic deletion variant of the Aspergillus fumigatus allergen Aspf1 displaying reduced IgE binding

• Published in: The FEBS journal 2005-05, Vol.272 (10), p.2536-2544
• Main Authors: Garciá‐Ortega, Lucía, Lacadena, Javier, Villalba, Mayte, Rodríguez, Rosalía, Crespo, Jesús F., Rodríguez, Julia, Pascual, Cristina, Olmo, Nieves, Oñaderra, Mercedes, del Pozo, Álvaro Martínez, Gavilanes, José G.
• Format: Article
• Language: English
• Subjects: allergen; Allergens - chemistry; Allergens - genetics; Allergens - immunology; Allergies; Amino Acid Sequence; Antigens, Plant; Aspergillus fumigatus; Aspergillus fumigatus - genetics; Aspergillus fumigatus - immunology; Aspf1; Binding sites; Cell Line, Tumor; Endoribonucleases - chemistry; Endoribonucleases - genetics; Endoribonucleases - immunology; Fungal Proteins - chemistry; Fungal Proteins - genetics; Fungal Proteins - immunology; Fungi; Humans; Immunoglobulin E - metabolism; Models, Molecular; Molecular Sequence Data; Protein Binding; Protein Structure, Secondary; Protein Synthesis Inhibitors - chemistry; Protein Synthesis Inhibitors - immunology; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - immunology; ribonuclease; ribotoxin; Sequence Alignment; Toxicity; α‐sarcin
• ISSN: 1742-464X; 1742-4658
• DOI: 10.1111/j.1742-4658.2005.04674.x

Aspergillus fumigatus is responsible for many allergic respiratory diseases, the most notable of which − due to its severity − is allergic bronchopulmonary aspergillosis. Aspf1 is a major allergen of this fungus: this 149‐amino acid protein belongs to the ribotoxin family, whose best characterized member is α‐sarcin (EC 3.1.27.10). The proteins of this group are cytotoxic ribonucleases that degrade a unique bond in ribosomal RNA impairing protein biosynthesis. Aspf1 and its deletion mutant Aspf1Δ(7–22) have been produced as recombinant proteins; the deleted region corresponds to an exposed β‐hairpin. The conformation of these two proteins has been studied by CD and fluorescence spectroscopy. Their enzymatic activity and cytotoxicity against human rhabdomyosarcoma cells was also measured and their allergenic properties have been studied by using 58 individual sera of patients sensitized to Aspergillus. Aspf1Δ(7–22) lacks cytotoxicity and shows a remarkably reduced IgE reactivity. From these studies it can be concluded that the deleted β‐hairpin is involved in ribosome recognition and is a significant allergenic region.