Characterization of a novel allergen, a major IgE-binding protein from Aspergillus flavus, as an alkaline serine protease.
Aspergillus species of fungi have been known to be one of the most prevalent aeroallergens. One important A. flavus allergen (Asp fl 1) was identified by means of immunoblotting with a serum pool of allergic patients on a two-dimensional electrophoretic gel. The cDNA coding for Asp fl 1 was cloned and sequenced. The clone encodes a full-length protein of 403 amino acid precursors of 42 kDa. After cleavage of a putative signal peptide of 21 amino acids and a prepeptide of 100 amino acids, a mature protein of 282 amino acids was obtained with a molecular mass of 33 kDa and a pI of 6.3. A degree of identity was found in a range of 27 to 84% among related allergens derived from bacteria allergen subtilisin, mold allergen Pen c 1, and virulence factor of A. fumigatus. Recombinant Asp fl 1 (rAsp fl 1) was cloned into vector pQE-30 and expressed in E. coli M15 as a histidine-tag fusion protein and purified to homogeneity. The IgE binding capacity of rAsp fl 1 was tested by immunoblotting using a serum pool of Aspergillus-allergic patients. Recombinant allergen cross-reacted strongly with IgE specific for natural Asp fl 1 and Pen c 1, indicating that common IgE epitopes may exist between allergens of A. flavus and P. citrinum.
Yu
C J
CJ
Institute of Biochemistry, College of Medicine, National Taiwan University, Institute of Biological Chemistry, Academia, Taipei, Taiwan.
Chiou
S H
SH
Lai
W Y
WY
Chiang
B L
BL
Chow
L P
LP
D000485:Allergens / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D000595:Amino Acid Sequence; D001231:Aspergillus flavus / Q000276:immunology*; D001483:Base Sequence; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D017385:Sequence Homology; D012697:Serine Endopeptidases / Q000032:analysis* / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*
D000485:Allergens; C120961:Asp fl 1 allergen; D007073:Immunoglobulin E; D012697:Serine Endopeptidases