Mapping of IgE binding regions in the major rat urinary protein, alpha 2u-globulin, using overlapping peptides.
Exposure to laboratory animals poses a hazard for development of occupational allergy. Identification of antigenic determinants of allergenic proteins may be valuable for immunotherapeutic purposes. Overlapping peptides of the major allergen in rat urine, Rat n 1.02, corresponding to the protein alpha2u-globulin were synthesised on solid support and screened simultaneously to locate IgE binding linear epitopes using a simple modified ELISA procedure. Thirty-nine peptides were synthesised, each 8 amino acids long with 4 amino acids overlaps. Sera from fifteen rat-sensitized subjects were analyzed and as controls sera from 7 non-rat-sensitized individuals were used. In general low binding and a great individual variation between sera from rat allergic individuals were seen. Some peptides were more frequently recognized by IgE antibodies in sera from rat allergics. These peptides were mainly clustered towards the N-terminal and C-terminal parts of the protein. Taken together our data suggest the existence of linear IgE binding epitopes in the rat urine allergen, Rat n 1.02. However, the role of these sequences in the allergic reaction needs further investigation.
Bayard
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Department of Occupational Medicine, National Institute for Working Life, Solna, Sweden. bayard@niwl.se
Siddique
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Berzins
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Troye-Blomberg
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Hellman
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U
Vesterberg
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D000510:Alpha-Globulins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000032:analysis / Q000276:immunology*; D006801:Humans; D006967:Hypersensitivity / Q000097:blood / Q000276:immunology; D007073:Immunoglobulin E / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism; D011506:Proteins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D017386:Sequence Homology, Amino Acid; D012984:Software
D000510:Alpha-Globulins; D018985:Epitopes, B-Lymphocyte; D010446:Peptide Fragments; D011506:Proteins; C013803:alpha 2u globulin; C035737:major urinary proteins; D007073:Immunoglobulin E