title: Structural and Functional Characterization of the Major Allergen Amb a 11 from Short Ragweed Pollen.
abstract: Allergy to the short ragweed (Ambrosia artemisiifolia) pollen is a major health problem. The ragweed allergen repertoire has been recently expanded with the identification of Amb a 11, a new major allergen belonging to the cysteine protease family. To better characterize Amb a 11, a recombinant proform of the molecule with a preserved active site was produced in Escherichia coli, refolded, and processed in vitro into a mature enzyme. The enzymatic activity is revealed by maturation following an autocatalytic processing resulting in the cleavage of both N- and C-terminal propeptides. The 2.05-Å resolution crystal structure of pro-Amb a 11 shows an overall typical C1A cysteine protease fold with a network of molecular interactions between the N-terminal propeptide and the catalytic triad of the enzyme. The allergenicity of Amb a 11 was confirmed in a murine sensitization model, resulting in airway inflammation, production of serum IgEs, and induction of Th2 immune responses. Of note, inflammatory responses were higher with the mature form, demonstrating that the cysteine protease activity critically contributes to the allergenicity of the molecule. Collectively, our results clearly demonstrate that Amb a 11 is a bona fide cysteine protease exhibiting a strong allergenicity. As such, it should be considered as an important molecule for diagnosis and immunotherapy of ragweed pollen allergy.
authors.affiliation: From Research and Development, Stallergenes Greer, 92160 Antony, France.
authors.forename: Rachel
authors.initials: R
authors.lastname: Groeme
authors.affiliation: From Research and Development, Stallergenes Greer, 92160 Antony, France.
authors.forename: Sabi
authors.initials: S
authors.lastname: Airouche
authors.affiliation: the Department of Protein Biochemistry and Proteomics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Šlechtitelů 27, CZ-78371 Olomouc, Czech Republic.
authors.forename: David
authors.initials: D
authors.lastname: Kopečný
authors.affiliation: From Research and Development, Stallergenes Greer, 92160 Antony, France.
authors.forename: Judith
authors.initials: J
authors.lastname: Jaekel
authors.affiliation: the SOLEIL Synchrotron, PROXIMA 2A, Saint Aubin-BP 48, 91192 Gif sur Yvette Cedex, France.
authors.forename: Martin
authors.initials: M
authors.lastname: Savko
authors.affiliation: From Research and Development, Stallergenes Greer, 92160 Antony, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Berjont
authors.affiliation: From Research and Development, Stallergenes Greer, 92160 Antony, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Bussieres
authors.affiliation: From Research and Development, Stallergenes Greer, 92160 Antony, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Le Mignon
authors.affiliation: the Institut Jean-Pierre Bourgin, Institut National de la Recherche Agronomique, AgroParisTech, Route de St-Cyr, 78026 Versailles, France, and.
authors.forename: Franck
authors.initials: F
authors.lastname: Jagic
authors.affiliation: the Vienna Challenge Chamber, Allergy Center Vienna West, A-1150 Vienna, Austria.
authors.forename: Petra
authors.initials: P
authors.lastname: Zieglmayer
authors.affiliation: From Research and Development, Stallergenes Greer, 92160 Antony, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Baron-Bodo
authors.affiliation: From Research and Development, Stallergenes Greer, 92160 Antony, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Bordas-Le Floch
authors.affiliation: From Research and Development, Stallergenes Greer, 92160 Antony, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Mascarell
authors.affiliation: the Institut Jean-Pierre Bourgin, Institut National de la Recherche Agronomique, AgroParisTech, Route de St-Cyr, 78026 Versailles, France, and.
authors.forename: Pierre
authors.initials: P
authors.lastname: Briozzo
authors.affiliation: From Research and Development, Stallergenes Greer, 92160 Antony, France, pmoingeon@stallergenes.com.
authors.forename: Philippe
authors.initials: P
authors.lastname: Moingeon
mesh_terms: D000485:Allergens / Q000737:chemistry / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D052179:Antigens, Plant / Q000276:immunology*; D020134:Catalytic Domain; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D057056:Cysteine Proteases / Q000737:chemistry* / Q000276:immunology; D004792:Enzyme Precursors / Q000737:chemistry* / Q000276:immunology; D005260:Female; D006860:Hydrogen Bonding; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010936:Plant Extracts / Q000276:immunology*; D010940:Plant Proteins / Q000737:chemistry* / Q000276:immunology; D011499:Protein Processing, Post-Translational; D059748:Proteolysis; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology* / Q000517:prevention & control
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D004792:Enzyme Precursors; D010936:Plant Extracts; D010940:Plant Proteins; C433085:ragweed pollen; D057056:Cysteine Proteases
keywords: allergen; cysteine protease; immunotherapy; protein processing; ragweed; structure-function