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J. Biol. Chem., Vol. 277, Issue 26, 23684-23692, June 28, 2002
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From the Bet v 1 is a 17-kDa protein abundantly present in
the pollen of the White birch tree and is the primary cause of birch
pollen allergy in humans. Its three-dimensional structure is remarkable in that a solvent-accessible cavity traverses the core of the molecule.
The biological function of Bet v 1 is unknown, although it is
homologous to a family of pathogenesis-related proteins in plants. In
this study we first show that Bet v 1 in the native state is able to
bind the fluorescent probe 8-anilino-1-naphthalenesulfonic acid (ANS).
ANS binds to Bet v 1 with 1:1 stoichiometry, and NMR data indicate that
binding takes place in the cavity. Using an ANS displacement assay, we
then identify a range of physiologically relevant ligands, including
fatty acids, flavonoids, and cytokinins, which generally bind with low
micromolar affinity. The ability of these ligands to displace ANS
suggests that they also bind in the cavity, although the exact binding
sites seem to vary among different ligands. The cytokinins, for
example, seem to bind at a separate site close to ANS, because they
increase the fluorescence of the ANS·Bet v 1 complex. Also,
the fluorescent sterol dehydroergosterol binds to Bet v 1 as
demonstrated by direct titrations. This study provides the first
qualitative and quantitative data on the ligand binding properties of
this important pollen allergen. Our findings indicate that ligand
binding is important for the biological function of Bet v 1.
The Major Birch Allergen, Bet v 1, Shows Affinity for a Broad
Spectrum of Physiological Ligands*
,,¶
Department of Biotechnology, Institute of
Life Sciences, Aalborg University, Sohngaardsholmsvej 49, Aalborg
DK-9000 and the § Biochemical Allergy Research,
ALK-Abelló A/S, Bøge Allé 6-8, Hørsholm
DK-2970, Denmark
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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