Abstract
Among proteins separated from an extract of Camellia japonica pollen by 2D-PAGE, three proteins, Cj 57, Cj 53 and Cj 50 with molecular weights of 57, 53 and 50kDa respectively, were detected as allergens by Western blotting using serum from a patient with camellia pollinosis. Moreover, the protein Cj 50 was identified as an enolase by mass spectrometry, and the amino acid sequence deduced by cloning of the Cj 50 gene confirmed that the protein is an enolase. Furthermore, the protein partially purified by column chromatography was found to have enolase activity. An antibody rised against Cj 50 recognized Cj 53 as well as Cj 50. Cj 53 may be an isoform of Cj 50 because it always was detected in the enolase fraction with Cj 50. These proteins were detected in the extracts of pollen grain, flower and leaf by Western blotting using the antibody, but Cj 53 was detected only in the pollen grain. Immuno-electron microscopy detected the labels of the antibody in the cytosol and generative cell nucleus of pollen, and in the cell nucleus of leaf cells. The immuno-labeling in the nucleus suggests that Cj 50 and/or Cj 53 have another physiological function besides enolase activity.
