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.

From Research and Development Stallergenes Greer 92160 Antony France

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

the Institut Jean Pierre Bourgin Institut National de la Recherche Agronomique AgroParisTech Route de St Cyr 78026 Versailles France and

the SOLEIL Synchrotron PROXIMA 2A Saint Aubin BP 48 91192 Gif sur Yvette Cedex France

the Vienna Challenge Chamber Allergy Center Vienna West A 1150 Vienna Austria

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PDB
1PCI

PDB
1S4V

PDB
1XKG

PDB
2AS8

PDB
3TNX

PDB
5EF4

PDB
5EGW

PDB
7PCK

PDB
9PAP