Characterization of high-molecular-mass allergens in oilseed rape pollen.
BACKGROUND
Oilseed rape pollen allergies have been previously described as the result of cross-sensitization with various pollens. Recently, several proteins have been identified as oilseed rape allergens. The aim of the present work was the characterization of oilseed rape pollen allergens by two-dimensional (2-D) gel analysis and amino acid microsequencing.


METHODS
Water extractable proteins from oilseed rape pollen were separated by isoelectrofocusing and then transferred onto a nitrocellulose sheet. Twenty-one human sera from pollen- or mustard-allergic individuals were screened for their reactivity to oilseed rape proteins. Eleven sera possessed IgE which recognized oilseed rape pollen proteins and one serum was selected for further 2-D characterization and amino acid microsequencing of the allergens.


RESULTS
The results showed that three molecules from oilseed rape pollen were identified as oilseed rape allergens which have not yet been described. These three proteins were molecules of 70 kD with a pI >8, 40 kD with a pI around 10 and 80 kD with a pI around 5. These proteins displayed identities with the berberine bridge protein, a receptor-like protein kinase and the cobalamin-independent methionine synthetase from Arabidopsis thaliana, respectively. The genes encoding the putative Arabidopsis molecules are located on chromosome 1 (berberine bridge protein) and chromosomes 3 and 4 (receptor-like protein kinases).


CONCLUSIONS
These results show that certain high-molecular-mass proteins from oilseed rape pollen are allergens.
Chardin
H
H
Institut Pasteur, Unité d'Immuno-Allergie, Paris, France. hchardin@pasteur.fr
Mayer
C
C
Sénéchal
H
H
Tepfer
M
M
Desvaux
F X
FX
Peltre
G
G
D013766:5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D015153:Blotting, Western; D001937:Brassica / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D011058:Pollen / Q000737:chemistry* / Q000276:immunology; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D011852:Radioallergosorbent Test; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000485:Allergens; D010940:Plant Proteins; D007073:Immunoglobulin E; D013766:5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase; D011494:Protein Kinases