title: Ribonuclease activity dependent cytotoxicity of Asp fl, a major allergen of A. fumigatus.
abstract: A major allergen/antigen, Asp fl, secreted by Aspergillus fumigatus exhibits cytotoxicity towards eukaryotic cell lines. Asp fl inhibited protein synthesis in RAW cells with an IC50 of 4.5 nM and also degraded ribosomal RNA of RAW cells at a similar concentration. Ribosomal inactivation by Asp fl may be the probable mechanism for protein synthesis inhibition. Specific ribonuclease activity of Asp fl was observed to be 100,000 U/mg. Presence of strong RNase activity in Asp fl was further confirmed by agar gels containing yeast RNA. Electrophoretic run on agarose gels showed that Asp fl degrades all species of naked RNA. Modification of histidine residues of Asp fl with diethyl pyrocarbonate and alkylation of cysteines with iodoacetamide resulted in loss of ribonuclease activity and cytotoxicity of Asp fl. The current study establishes the ribonuclease activity of a purified major allergen of A. fumigatus that inhibits protein synthesis and kills the eukaryotic cells.
authors.affiliation: Centre for Biochemical Technology, Delhi, India.
authors.forename: T
authors.initials: T
authors.lastname: Madan
authors.forename: N
authors.initials: N
authors.lastname: Arora
authors.forename: P U
authors.initials: PU
authors.lastname: Sarma
mesh_terms: D000485:Allergens / Q000276:immunology / Q000378:metabolism*; D000946:Antigens, Fungal / Q000276:immunology / Q000378:metabolism*; D052179:Antigens, Plant; D001232:Aspergillus fumigatus / Q000276:immunology*; D002460:Cell Line; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic; D003603:Cytotoxins / Q000276:immunology / Q000378:metabolism*; D004047:Diethyl Pyrocarbonate; D004587:Electrophoresis, Agar Gel; D004791:Enzyme Inhibitors; D004797:Enzyme-Linked Immunosorbent Assay; D005656:Fungal Proteins / Q000276:immunology / Q000378:metabolism*; D007460:Iodoacetamide; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000276:immunology / Q000378:metabolism*; D012331:RNA, Fungal / Q000378:metabolism; D012335:RNA, Ribosomal / Q000378:metabolism; D012260:Ribonucleases / Q000276:immunology / Q000378:metabolism*
chemical_list: C081947:ASPF1 protein, Aspergillus fumigatus; D000485:Allergens; D000946:Antigens, Fungal; D052179:Antigens, Plant; D003603:Cytotoxins; D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D011500:Protein Synthesis Inhibitors; D012331:RNA, Fungal; D012335:RNA, Ribosomal; C036196:MITF protein, Aspergillus restrictus; D012260:Ribonucleases; D004047:Diethyl Pyrocarbonate; D007460:Iodoacetamide