Molecular cloning and allergenicity of Pen j 1, a major allergen of kuruma prawn, Penaeus japonicus.
Tropomyosins have been identified as a common allergen in crustaceans, but their allergenicity is not well understood. In the present study, we isolated an allergen, Pen j 1, a tropomyosin from kuruma prawn Penaeus japonicus, and determined its N-terminal amino acid sequence. The cDNA encoding the allergen was cloned by 5'- and 3'-rapid amplification of cDNA ends (RACE), and was found to code for a protein which consists of 284 amino acid residues. Sequencing analyses indicated for the first time that mature tropomyosin is formed by the elimination of a leader peptide of nine amino acid residues. To elucidate the binding sites of IgE antibodies in the sera of shrimp-sensitive patients, various recombinant peptides were expressed in Escherichia coli as fusion proteins with glutathione S-transferase (GST), and the examined with regard to reactivity with IgE antibodies. The IgE-binding epitopes were found to locate over the whole sequence of the allergen, and the IgE antibodies in the sera were found to recognize strongly its C-terminal region.
Kunimoto
Ayumi
A
Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Kuboki, Soja, Okayama 719-1197, Japan.
Sisino
Takako
T
Sakai
Kentaro
K
Matsumoto
Tomoaki
T
Takahashi
Kyoko
K
Yamashita
Hiromi
H
Hiemori
Miki
M
Tsuji
Hideaki
H
Kimoto
Masumi
M
D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D005512:Food Hypersensitivity / Q000276:immunology; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology / Q000378:metabolism; D007223:Infant; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D010446:Peptide Fragments / Q000235:genetics / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D014335:Tropomyosin / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification
D000485:Allergens; D018076:DNA, Complementary; D010446:Peptide Fragments; D014335:Tropomyosin; D007073:Immunoglobulin E