Reduction of antigenicity of Cry j 1, a major allergen of Japanese cedar pollen, by thermal denaturation.
The soluble aggregates of Cry j 1, a major allergen of Japanese cedar pollen, were formed without any coagulates during heat treatment at acidic pH 5, as shown in HPLC and SDS-PAGE patterns. A remarkable change in the CD spectrum was observed between native and heat-denatured Cry j 1 at a linear rate of 1 degrees C/min from 40 to 90 degrees C. The negative peak of native Cry j 1 at 222 nm was moved to 218 nm, suggesting the transition of an alpha-helix to beta-structure during heat denaturation. The increase in beta-structure was also observed during heat denaturation by monitoring the fluorescence with Thioflavin T. These results suggest that Cry j 1 forms intermolecular cross-beta-structure between denatured proteins during heating at 90 degrees C. The antigenicity of Cry j 1 detected by dot-blotting was greatly diminished during heating at a linear rate of 1 degrees C/min from 40 to 90 degrees C without any coagulates. These results suggest that IgE epitopes exposed on the molecular surface of Cry j 1 was buried inside soluble aggregates through intermolecular beta-structure formed by heating.
Department of Biological Chemistry, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
Rieko
R
Aoki
Akira
A
Saito
Masakatsu
M
Usui
Hiroyuki
H
Azakami
Akio
A
Kato
D000485:Allergens / Q000737:chemistry* / Q000276:immunology; D052179:Antigens, Plant / Q000737:chemistry* / Q000276:immunology; D032485:Cryptomeria / Q000737:chemistry / Q000276:immunology*; D006358:Hot Temperature; D011058:Pollen / Q000737:chemistry* / Q000276:immunology; D011489:Protein Denaturation; D017433:Protein Structure, Secondary
D000485:Allergens; D052179:Antigens, Plant