Cloning, expression and characterization of the major latex allergen prohevein
- 1 November 1998
- journal article
- research article
- Published by Wiley in Clinical and Experimental Allergy
- Vol. 28 (11) , 1418-1426
- https://doi.org/10.1046/j.1365-2222.1998.00422.x
Abstract
About 70–80% of latex allergic health care workers are sensitized to prohevein (Hev b 6.01), a 20 kDa cysteine-rich chitin-binding protein of Hevea latex. This study reports on the bacterial cloning, expression and immunochemical characterization of rHev b 6.01. Prohevein was expressed in the periplasmatic space of Escherichia coli as maltose binding protein (MBP) fusion protein and purified to homogeneity after factor Xa cleavage. The IgE binding capacity of both rHev b 6.01 and prohevein isolated from fresh Hevea latex was compared by immunoblotting experiments using sera of latex-allergic patients. The diagnostic value of rHev b 6.01 was analysed by enzyme allergosorbent test (EAST). Two different cDNA clones of rHev b 6.01 were established. The deduced amino acid sequence of both clones revealed two and three amino acid differences in the C-terminal domain of prohevein compared with the original database entry. Purified rHev b 6.01 bound with high affinity to chitin as its natural counterpart isolated from natural latex. In IgE-immunblotting using sera of affected subjects binding intensity to both proteins was comparable indicating a very high antigenic similarity. The diagnostic value of MBP-prohevein was tested in EAST using sera of 33 latex-allergic subjects. The in vitro test showed high sensitivity and specificity and proved the diagnostic value of uncleaved MBP-prohevein. The production of recombinant latex key allergens with defined quality like prohevein is a straightforward strategy for the development of standardized in vitro test systems.Keywords
This publication has 21 references indexed in Scilit:
- Diagnosis of latex allergyAllergy, 1997
- Latex allergen databaseElectrophoresis, 1997
- Latex allergy diagnosis: in vivo and in vitro standardization of a natural rubber latex extractAllergy, 1997
- Identification, Cloning, and Sequence of a Major Allergen (Hev b 5) from Natural Rubber Latex (Hevea brasiliensis)Journal of Biological Chemistry, 1996
- A Novel Acidic Allergen, Hev b 5, in LatexJournal of Biological Chemistry, 1996
- Natural rubber latex allergyAllergy, 1996
- Prohevein from the rubber tree (Hevea brasiliensis) is a major latex allergenClinical and Experimental Allergy, 1995
- CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choiceNucleic Acids Research, 1994
- Vectors that facilitate the expression and purification of foreign peptides in Escherichia coli by fusion to maltose-binding proteinGene, 1988
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970