[Immunological analysis and mass spectrometry identification of the major allergen from Cladosporium cladosporioides].
OBJECTIVE
To analyze and identify the major allergen from Cladosporium cladosporioides.


METHODS
The Cladosporium cladosporioides were collected by means of glass dish exposed by air. After they were cultured, the protein of the Cladosporium cladosporioides was extracted with carbonate buffer solution. The major allergen in the Cladosporium cladosporioides was characterized by Western-blotting after SDS-PAGE. After in-gel digestion, the peptide mass values of the major allergen which were obtained by matrix-assisted laxer desorption ionization time-of-flight mass spectrometry, (MALDI-TOF-MS) were used as inputs to search corresponding proteins with the Mascot (http://www. matrixscience. corn).


RESULTS
There were fourteen protein bands visible on the SDS-PAGE gel in the crude extracts. The most abundant ten bands were 72 kD, 61 kD, 42 kD, 36 kD, 35 kD, 34 kD, 24 kD, 22 kD, 18 kD and 14 kD proteins, respectively. The protein whose molecular mass was 18 kD was identified by Western-blotting as major allergen. The major allergen was similar to the 18 kD ribosomal protein of Psychrobacter arcticum based on peptide mass fingerprint analysis (MALDI-TOF-MS).


CONCLUSIONS
The major allergen from Cladosporium cladosporioides, a ribosomal protein about 18 kD, was analyzed and identified.
Li
Li
L
Allergy and Immunology Institute, Shenzhen University, Shenzhen 518060, China. ylili@szu.edu.cn
Li
Qiyuan
Q
Liu
Zhigang
Z
Wu
Haiqiang
H
D000485:Allergens / Q000032:analysis / Q000276:immunology*; D001426:Bacterial Proteins / Q000032:analysis / Q000276:immunology*; D002963:Cladosporium / Q000737:chemistry / Q000276:immunology*; D012269:Ribosomal Proteins / Q000032:analysis / Q000276:immunology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D000485:Allergens; D001426:Bacterial Proteins; D012269:Ribosomal Proteins