Production of recombinant mite allergen Der fI in insect cells and characterization of products--removal of pro-sequence is essential to IgE-binding activity.
Der fI is a major mite proteinaceous allergen found in house dust. We produced recombinant Der fI (reDer fI) in insect cells using a baculovirus expression system. Based on the molecular mass and N-terminal amino acid (aa) sequence analysis, reDer fI was found to be compromised of a mixture of two proteins, each of which includes a pro-sequence of different length. The reDer fI had IgE-binding activity at only 20% of that of native Der fI. The removal of the pro-sequence in an acidic solution drastically increased IgE-binding activity to almost the same as that of native Der fI, showing that the presence of the pro-sequence is inhibitory to the IgE-binding activity of Der fI.
Shoji
H
H
Institute for Production Research and Development, Nikka Whisky Distilling Co., Ltd., Chiba, Japan.
Hanawa
M
M
Shibuya
I
I
Hirai
M
M
Yasuhara
T
T
Okumura
Y
Y
Yamakawa
H
H
D000485:Allergens / Q000096:biosynthesis / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D039741:Antigens, Dermatophagoides; D016367:Baculoviridae / Q000235:genetics*; D002478:Cells, Cultured; D005822:Genetic Vectors; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D007073:Immunoglobulin E / Q000378:metabolism*; D008925:Mites; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis; D018411:Spodoptera / Q000166:cytology
D000485:Allergens; D039741:Antigens, Dermatophagoides; D006023:Glycoproteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E