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Originally published In Press as doi:10.1074/jbc.M002854200 on April 27, 2000

J. Biol. Chem., Vol. 275, Issue 28, 21572-21577, July 14, 2000
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Crystal Structure of the Allergen Equ c 1
A DIMERIC LIPOCALIN WITH RESTRICTED IgE-REACTIVE EPITOPES*

Marie-Bernard LascombeDagger , Christophe Grégoire§, Pascal Poncet§, Gisele A. TavaresDagger ||, Isabelle Rosinski-Chupin**, Jacques Rabillon§, Hany Goubran-Botros§, Jean-Claude MaziéDagger Dagger , Bernard David§, and Pedro M. AlzariDagger §§

From the Dagger  Unité de Biochimie Structurale (CNRS URA 2185), § Unité d'Immuno-Allergie, ** Unité de Génétique et de Biochimie du Développement, and Dagger Dagger  Laboratoire d'Ingénierie des Anticorps, Institut Pasteur, 25 et 28 rue du Docteur Roux, 75724 Paris Cedex 15, France

The three-dimensional structure of the major horse allergen Equ c 1 has been determined at 2.3 Å resolution by x-ray crystallography. Equ c 1 displays the typical fold of lipocalins, a beta -barrel flanked by a C-terminal alpha -helix. The space between the two beta -sheets of the barrel defines an internal cavity that could serve, as in other lipocalins, for the binding and transport of small hydrophobic ligands. Equ c 1 crystallizes in a novel dimeric form, which is distinct from that observed in other lipocalin dimers and corresponds to the functional form of the allergen. Binding studies of point mutants of the allergen with specific monoclonal antibodies raised in mouse and IgE serum from horse allergic patients allowed to identify putative B cell antigenic determinants. In addition, total inhibition of IgE serum recognition by a single specific monoclonal antibody revealed the restricted nature of the IgE binding target on the molecular surface of Equ c 1.

* This work was supported by grants from the Institut Pasteur and CNRS (France) and the European Union for the TMR/LSF program to the EMBL Hamburg Outstation, reference number ERBFMGECT980134.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.

Present address: University of Virginia, Dept. of Medicine, Box 225, Health Sciences Center, Charlottesville, VA 22908.

|| Present address: Dept. of Molecular Biophysics & Biochemistry, Yale University, 260 Whitney Ave., New Haven, CT 06511.

§§ To whom correspondence should be addressed: Unité de Biochimie Structurale, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France. Tel.: (33) 1 45 68 86 07; Fax: (33) 1 45 68 86 04; E-mail: alzari@pasteur.fr.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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