Cloning of the patatin-like latex allergen Hev b 7, its expression in the yeast Pichia pastoris and its immunological characterization.
The 43-kD latex allergen Hev b 7 was purified from the latex of Hevea brasiliensis and identified by N-terminal and internal peptide sequences as highly homologous to patatins. Patatins are storage proteins encoded by a multigene family found in plants such as potato and tomato. We have obtained a cDNA clone coding for a cytoplasmic form of Hev b 7. The recombinant protein was expressed in the methylotrophic yeast Pichia pastoris at 10 mg/l culture supernatant. Both natural Hev b 7 and rHev b 7 were recognized by IgE in 11% of the latex-allergic patients. rHev b 7 inhibited binding to its counterpart in natural rubber latex extracts. Purified rHev b 7 used at concentrations of 10 micrograms/ml in skin prick tests produced wheal-and-flare reactions of sizes equal to those produced by nHev b 7. Furthermore, we were able to show that rHev b 7 possessed esterase activity. A plant expression system for the production of larger quantities of recombinant latex allergens as an alternative to the preparation from H. brasiliensis sap is discussed.
Breiteneder
H
H
Department of General and Experimental Pathology, University of Vienna, Vienna, Austria. Heimo.Breiteneder@akh-wien.ac.at
Sowka
S
S
Wagner
S
S
Krebitz
M
M
Hafner
C
C
Kinaciyan
T
T
Yeang
H Y
HY
Scheiner
O
O
D000485:Allergens / Q000235:genetics / Q000276:immunology*; D052179:Antigens, Plant; D002265:Carboxylic Ester Hydrolases; D019665:Euphorbiaceae; D020315:Latex Hypersensitivity; D010843:Pichia / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D000485:Allergens; D052179:Antigens, Plant; D010940:Plant Proteins; D011994:Recombinant Proteins; C113002:allergen Hev b 7; C043222:patatin protein, Solanum tuberosum; D002265:Carboxylic Ester Hydrolases