TY - JOUR
T1 - Insights into a conformational epitope of Hev b 6.02 (hevein)
AU - Reyes-López, César A.
AU - Hernández-Santoyo, Alejandra
AU - Pedraza-Escalona, Martha
AU - Mendoza, Guillermo
AU - Hernández-Arana, Andrés
AU - Rodríguez-Romero, Adela
N1 - Funding Information:
We thank Peter Kuhn for beamline support. This work is based on research conducted at SSRL, which is funded by the Department of Energy, Office of Basic Energy Sciences. The Biotechnology Program is supported by the National Institutes of Health, National Center for Research Resources, Biomedical Technology Program and the Department of Energy. Preliminary X-ray experiments were done at LUEP IQ-UNAM. We thank Dr. Edgar Zenteno and Dra. Concepción Agundis (School of Medicine-UNAM) for their help with the production of antibodies and CONACYT (Grant 32417-E).
PY - 2004/1/30
Y1 - 2004/1/30
N2 - Hevein (Hev b 6.02) is a major IgE-binding allergen in natural rubber latex and manufactured products. Both tryptophans (Trp21 and Trp 23) of the hevein molecule were chemically modified with BNPS-skatole (2-nitrophenylsulfenyl-3-methyl-3′-bromoindolenine); derivatized allergen failed to significantly inhibit binding of serum IgE in ELISA assays. Similarly, skin prick tests showed that hevein-positive patients gave no response with the modified allergen. Dot blot experiments carried out with anti-hevein mono- and polyclonal antibodies confirmed the importance of Trp21 and Trp23 for antibody-recognition, and demonstrated the specific cross-reactivity of other molecules containing hevein-like domains. We also report the structure of Hev b 6.02 at an extended resolution (1.5Å) and compare its surface properties around Trp residues with those of similar regions in other allergens. Overall our results indicate that the central part of the protein, which comprises three aromatic and other acidic and polar residues, constitutes a conformational epitope.
AB - Hevein (Hev b 6.02) is a major IgE-binding allergen in natural rubber latex and manufactured products. Both tryptophans (Trp21 and Trp 23) of the hevein molecule were chemically modified with BNPS-skatole (2-nitrophenylsulfenyl-3-methyl-3′-bromoindolenine); derivatized allergen failed to significantly inhibit binding of serum IgE in ELISA assays. Similarly, skin prick tests showed that hevein-positive patients gave no response with the modified allergen. Dot blot experiments carried out with anti-hevein mono- and polyclonal antibodies confirmed the importance of Trp21 and Trp23 for antibody-recognition, and demonstrated the specific cross-reactivity of other molecules containing hevein-like domains. We also report the structure of Hev b 6.02 at an extended resolution (1.5Å) and compare its surface properties around Trp residues with those of similar regions in other allergens. Overall our results indicate that the central part of the protein, which comprises three aromatic and other acidic and polar residues, constitutes a conformational epitope.
KW - Allergens
KW - Hev b 6.02
KW - Hevein
KW - IgE-epitope
KW - Latex
KW - X-ray structure
UR - http://www.scopus.com/inward/record.url?scp=0346095164&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2003.12.068
DO - 10.1016/j.bbrc.2003.12.068
M3 - Artículo
SN - 0006-291X
VL - 314
SP - 123
EP - 130
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -