Purification and characterization of a novel isoallergen of a major Bermuda grass pollen allergen, Cyn d 1.
A novel immunoreactive isoallergen of a major Bermuda grass pollen allergen, Cyn d 1, was purified by the use of a combination of various chromatographic techniques, including high-performance liquid chromatography. This new isoallergen has a pI value of 9.1 and shows significant N-terminal sequence homology with other isoforms. Carbohydrate composition analysis revealed a 10.4% carbohydrate content consisting of 7 different sugar moieties, including arabinose, fucose, galactose, glucose, mannose, xylose and N-acetylglucosamine, as well as a trace amount of rhamnose. Upon periodate oxidation, the binding activities of the Cyn d 1 isoform to murine monoclonal antibodies and human serum IgE and IgG were reduced, suggesting the importance of the carbohydrate moiety in the immune response. The availability of the purified Cyn d 1 basic isoform will allow for further structural and immunological characterization, and ultimately for the design of an appropriate therapy.
Su
Song-Nan
SN
Department of Medical Research and Education, Veterans General Hospital-Taipei, Taipei, Taiwan, ROC. snsu@vghtpe.gov.tw
Peng
Ho-Jen
HJ
Yang
Sue-Yee
SY
Tsai
Lai-Chen
LC
Chow
Lu-Ping
LP
Huang
Shih-Wen
SW
D000485:Allergens / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis / Q000276:immunology; D052179:Antigens, Plant; D002241:Carbohydrates / Q000032:analysis / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera / Q000032:analysis / Q000276:immunology; D007073:Immunoglobulin E / Q000097:blood; D007074:Immunoglobulin G / Q000097:blood; D037102:Lectins / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D020033:Protein Isoforms / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification
D000485:Allergens; D000596:Amino Acids; D052179:Antigens, Plant; C071778:CYND1 protein, Cynodon dactylon; D002241:Carbohydrates; D007106:Immune Sera; D007074:Immunoglobulin G; D037102:Lectins; D010940:Plant Proteins; D020033:Protein Isoforms; D007073:Immunoglobulin E