Assignment of the disulfide bonds of Ole e 1, a major allergen of olive tree pollen involved in fertilization.
The most prevalent allergen from olive tree pollen, Ole e 1, consists of a single polymorphic polypeptide chain of 145 amino acids which includes six cysteine residues at positions 19, 22, 43, 78, 90 and 131. By using an homogeneous form of the allergen expressed in Pichia pastoris, the array of the disulfide bridges has been elucidated. Specific proteolysis with thermolysin and reverse-phase HPLC separation of the peptides allowed the determination of the disulfide bond between Cys43 and Cys78. Another thermolytic product, which contained three peptides linked by the remaining four cysteines, was digested with Glu-specific staphylococcal V8 protease and the products isolated by reverse-phase HPLC. Amino acid compositions and Edman degradation of the peptide products indicated the presence of the disulfide bonds at Cys19-Cys90 and Cys22-Cys131. These data can help in the analysis of the three-dimensional structure of the protein as well as in studies of its allergenic determinants.
González
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E
Departamento de Bioquímica y Biología Molecular, Facultad de Química, Universidad Complutense, Madrid, Spain.
Monsalve
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RI
Puente
X S
XS
Villalba
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M
Rodríguez
R
R
D000485:Allergens / Q000737:chemistry* / Q000235:genetics; D000596:Amino Acids / Q000032:analysis; D052179:Antigens, Plant; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004220:Disulfides / Q000737:chemistry*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics; D011058:Pollen; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013820:Thermolysin / Q000737:chemistry / Q000378:metabolism; D014197:Trees
D000485:Allergens; D000596:Amino Acids; D052179:Antigens, Plant; D004220:Disulfides; C081946:Ole e I protein, Olea europaea; D010940:Plant Proteins; D011994:Recombinant Proteins; D013820:Thermolysin