Two-dimensional IgE-binding spectrum of Japanese cedar (Cryptomeria japonica) pollen allergens.
BACKGROUND
Japanese cedar (Cryptomeria japonica) pollen is one of the most prevalent sources of the allergens that elicit rhinitis and conjunctivitis. Only Cry j 1 and Cry j 2 have been well characterized as the major allergens of this pollen.


OBJECTIVE
The aims of this study were to complete the repertoire of C. japonica pollen allergens, to investigate their variability with respect to IgE-reactive patterns and to identify the isoforms of Cry j 1 and Cry j 2 by proteome analysis.


METHODS
Proteins in C. japonica pollen separated on two-dimensional (2-D) polyacrylamide gel electrophoresis were immunodetected with IgE in sera of 40 subjects allergic to C. japonica pollen. Mass fingerprinting was used to elucidate the diversity of the major allergens.


RESULTS
2-D immunolabeling with individual patients' sera showed the distinguishable IgE-binding patterns inlaid with 4-87 spots from a total of 131 IgE-binding protein spots. At least 12 Cry j 1 (27.5-75% of IgE-binding frequency) and 3 Cry j 2 (32.5-40%) isoforms were localized. In total, 31 spots were found to be more reactive than the highest IgE-reactive isoform of Cry j 2.


CONCLUSIONS
The proteomics approaches showed great interindividual variation of IgE-binding patterns to C. japonica proteins and contributed to the repertoire of numerous C. japonica allergens other than Cry j 1 and Cry j 2. Protein microsequencing demonstrated more complicated multiplicity in Cry j 1 than previously known and new isoforms in Cry j 2.
Fujimura
Takashi
T
Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima, Japan.
Shigeta
Seiko
S
Kawamoto
Seiji
S
Aki
Tsunehiro
T
Masubuchi
Masanori
M
Hayashi
Takaharu
T
Yoshizato
Katsutoshi
K
Ono
Kazuhisa
K
D000485:Allergens / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D032485:Cryptomeria / Q000737:chemistry / Q000276:immunology*; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000737:chemistry / Q000276:immunology*; D008297:Male; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology*; D011058:Pollen / Q000737:chemistry / Q000276:immunology*; D020033:Protein Isoforms; D020539:Sequence Analysis, Protein; D021241:Spectrometry, Mass, Electrospray Ionization
D000485:Allergens; D052179:Antigens, Plant; C065290:Cry j I protein, Cryptomeria japonica; C065291:Cry j II protein, Cryptomeria japonica; D010940:Plant Proteins; D020033:Protein Isoforms; D007073:Immunoglobulin E