Identification of a polygalacturonase as a major allergen (Pla a 2) from Platanus acerifolia pollen Ignacio Ibarrola, PhD, M. Carmen Arilla, PhD, Alberto.

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Identification of a polygalacturonase as a major allergen (Pla a 2) from Platanus acerifolia pollen Ignacio Ibarrola, PhD, M. Carmen Arilla, PhD, Alberto Martínez, PhD, Juan A. Asturias, PhD Journal of Allergy and Clinical Immunology Volume 113, Issue 6, Pages 1185-1191 (June 2004) DOI: 10.1016/j.jaci.2004.02.031 Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 Electrophoretic analysis of purified Pla a 2. A, Coomassie-stained SDS-PAGE (a) and immunostaining with reactive sera (b) of fractions from the purification steps: raw extract (lanes 1), after the HiTrap SP column (lanes 2), and purified Pla a (lanes 3). B, Coomassie-stained agarose IEF (a) and IgE immunoblotting (b) of purified Pla a 2. C, Staining of purified Pla a 2 with the Glycan Detection Kit. As controls, Ole e 2 (lane 4) and Ole e 1 (lane 5) were used. Journal of Allergy and Clinical Immunology 2004 113, 1185-1191DOI: (10.1016/j.jaci.2004.02.031) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 SDS-PAGE immunoblotting inhibition of P acerifolia pollen extract. Serum pool of monosensitized patients preincubated with buffer alone (C) or different concentrations (in nanograms per milliliter) of purified Pla a 2 and BSA. Journal of Allergy and Clinical Immunology 2004 113, 1185-1191DOI: (10.1016/j.jaci.2004.02.031) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 ELISA-inhibition curves. The binding of specific human IgE to P acerifolia extract was inhibited by different concentrations of Pla a 2, P acerifolia extract, or BSA as a control. Journal of Allergy and Clinical Immunology 2004 113, 1185-1191DOI: (10.1016/j.jaci.2004.02.031) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 Sequence analysis of cDNA encoding Pla a 2: nucleotide and deduced amino acid sequences. Sequenced internal peptides are underlined, and predicted N-glycosylation sites are double underlined. The arrow indicates the experimental cleavage site for signal peptide. The nucleotide sequence for Pla a 2 has been deposited in the GenBank database under accession no. AJ586898. Journal of Allergy and Clinical Immunology 2004 113, 1185-1191DOI: (10.1016/j.jaci.2004.02.031) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 5 Pollen and fruit PG amino acid sequence alignment (accession nos. AJ586898, P24548, P35337, Q39786, P35339, P48979, P48978, Q02096, P35336, P05117, and P43212, respectively). Asterisks represent conserved residues. Amino acids were colored to illustrate conservation of physicochemical properties: yellow, cysteines; light green, hydrophobic; light blue, hydrophilic; dark blue, acidic; red, basic. Journal of Allergy and Clinical Immunology 2004 113, 1185-1191DOI: (10.1016/j.jaci.2004.02.031) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions