cDNA cloning of the 43-kDa latex allergen Hev b 7 with sequence similarity to patatins and its expression in the yeast Pichia pastoris.
IgE-mediated hypersensitivity to latex proteins present in health care products, particularly in latex gloves, has become an important public health problem in recent years. We purified natural Hev b 7, a 43-kDa patatin-like allergen from the latex of Hevea brasiliensis and determined several internal peptide sequences. A heterologous hybridization probe of a patatin gene of potato, to which these peptides could be aligned best, was used to screen a latex cDNA library. The cDNA encoded an acidic protein of 388 amino acids with a molecular mass of 42.9 kDa. The deduced amino acid sequence had 39-42% identity to patatins from Solanum tuberosum. The purified recombinant Hev b 7 expressed in the yeast Pichia pastoris displayed, similarly to patatins from S. tuberosum, esterase activity. Both natural and recombinant Hev b 7 were recognized by IgE from sera of latex-sensitized allergic individuals. In contrast to patatins from S. tuberosum and Nicotiana tabacum, natural Hev b 7 lacked an N-terminal leader peptide for targeting to the endoplasmatic reticulum and was not glycosylated. These results establish the 43-kDa patatin-like protein as a latex allergen and raise the possibility of different cellular localization and function compared to S. tuberosum patatins.
Sowka
S
S
Department of General and Experimental Pathology, University Hospital, Vienna, Austria.
Wagner
S
S
Krebitz
M
M
Arija-Mad-Arif
S
S
Yusof
F
F
Kinaciyan
T
T
Brehler
R
R
Scheiner
O
O
Breiteneder
H
H
D000485:Allergens / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D019665:Euphorbiaceae / Q000235:genetics*; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology*; D007840:Latex / Q000276:immunology*; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D011852:Radioallergosorbent Test; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D017386:Sequence Homology, Amino Acid
D000485:Allergens; D052179:Antigens, Plant; D018076:DNA, Complementary; D007840:Latex; D010940:Plant Proteins; D011994:Recombinant Proteins; C113002:allergen Hev b 7; C043222:patatin protein, Solanum tuberosum; D002265:Carboxylic Ester Hydrolases