Glycosylation site of the major allergen from olive tree pollen. Allergenic implications of the carbohydrate moiety.
The electrophoretic analysis of purified Ole e I, the major allergen from Olea europaea pollen, reveals the presence of two main variants, glycosylated (20.0 kDa) and non-glycosylated (18.5 kDa) components. The glycosylated variant has been identified as a concanavalin A-binding glycoprotein. Its carbohydrate moiety has a molecular mass of about 1.3 kDa (5% weight of the glycosylated allergen), based on mass spectrometry analysis. Enzymatic treatment of native Ole e I with the specific glycosidase PNGase F accounts for an oligosaccharide N-linked to the polypeptide chain. This treatment does not sensibly modify the secondary structure of the protein but diminishes the affinity of the allergen for specific IgE antibodies. Tryptic digestion of Ole e I reveals the presence of a single carbohydrate-containing peptide. This peptide was recognized by the sera of hypersensitive individuals. The amino acid sequence of this peptide is Phe-Lys-Leu-Asn-Thr-Val-Asn-Gly-Thr-Thr-Arg, asparagine at the seventh being the carbohydrate attaching site. The obtained data are discussed in terms of the potential role of the sugar moiety in the allergenic activity of Ole e I.
Batanero
E
E
Departamento de Bioquímica y Biología Molecular I, Facultad de Química, Universidad Complutense, Madrid, Spain.
Villalba
M
M
Rodríguez
R
R
D000485:Allergens / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D002241:Carbohydrates / Q000737:chemistry / Q000276:immunology*; D006031:Glycosylation; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011058:Pollen / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D017433:Protein Structure, Secondary; D014197:Trees / Q000276:immunology*
D000485:Allergens; D052179:Antigens, Plant; D002241:Carbohydrates; C081946:Ole e I protein, Olea europaea; D010940:Plant Proteins; D007073:Immunoglobulin E