Characterization of major allergens of Parietaria officinalis.
The major allergens of Parietaria officinalis were characterized with a panel of nine monoclonal antibodies (mAbs). The binding of mAbs and patients' IgE in Western blots revealed two proteins with similar molecular weights in the range of 8-10 kD. Analysis of the mAb-binding patterns in Western blots of P. officinalis extract under reducing and nonreducing conditions allows the mAbs to be divided into three different groups. mAbs of group I recognize the higher-molecular-weight component (9.4 kD), mAbs of group II recognize the lower component (8.8 kD) and mAbs of group III recognize both proteins. A comparable mAb-binding pattern was observed with Western blots of Parietaria judaica. The mAbs were used for affinity purification of the corresponding proteins from a P. officinalis extract. The purified proteins obtained with mAbs of group I-III inhibit the binding of patients' IgE (serum pool) to a high degree, indicating that they posses the major IgE-reactive epitopes. The affinity-purified proteins were subjected to SDS-PAGE, blotted and immunologically stained by mAb binding. The results confirmed those obtained with the complete extracts. The N-terminal amino acid sequences of the blotted proteins were analyzed. The sequences of all the proteins contained highly conserved regions: GGVV (positions 4-7) and MPPLL (positions 11-15), alternating with highly variable regions (positions 1-3 for group II and 8-10 for group I). A specific group I sequence appears to be at position 1-3 with the amino acids APA and a specific group II sequence appears to be at position 8-10 with the amino acids GAL. It is possible that the two similar proteins are isoforms of Par o 1.
Kahlert
H
H
Allergopharma Joachim Ganzer KG, Reinbek, Germany.
Weber
B
B
Teppke
M
M
Wahl
R
R
Cromwell
O
O
Fiebig
H
H
D000485:Allergens / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001666:Binding Sites, Antibody; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology*; D007073:Immunoglobulin E / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010940:Plant Proteins; D011485:Protein Binding; D012967:Sodium Dodecyl Sulfate
D000485:Allergens; D000911:Antibodies, Monoclonal; D006023:Glycoproteins; C083860:Par j I protein, Parietaria judaica; C072038:Par o I protein, Parietaria officinalis; D010940:Plant Proteins; D012967:Sodium Dodecyl Sulfate; D007073:Immunoglobulin E