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mnfr2033-sup-0001-FigureS1.docx1015K

Figure 1. Sequence identification and amino acid sequence alignment of Api g 6 and other nsLTP2. Tryptic peptides of Api g 6 were identified by MS and N-terminal sequencing and analyzed fragments are represented by solid and dotted lines, respectively (A). The mature sequences of Api g 6 (Apium graveolens; P86809) and nsLTP2s from olive (Olea europea; fruit; J7FQE3), birdsfoot trefoil (Lotus japonicus; tissue not defined; I3SYA6), common tobacco (Nicotiana tabacum; leaf trichomes; E3W9R1), potato (Solanum tuberosum; fruit; Q8H9B7), soybean (Glycine max; fruit; I1N5L5), kashgar tamarisk (Tamarix hispida; mixture of roots, stems and leaves; C0KHK5), apricot (Prunus armeniaca; fruit; P82353), black cottonwood (Populus trichocarpa; leaves; A9P8Y3), common grape wine (Vitis vinifera; genomic DNA; D7TSZ9), sesame (Sesamum indicum; seeds; A5JUZ7), barrel medic (Medicago truncatula; genomic DNA; B7FM99), blue gum (Eucalyptus globulus; tissue not defined; I0IK56), mouse-ear cress (Arabidopsis thaliana; tissue not defined; Q9LJQ3), castor oil (Ricinus communis; tissue not defined; B9SM06), cowpea (Vigna unguiculata; root hairs; Q43681), groundsel (Senecio odorus; tissue not defined; Q41378), Asian rice (Oryza sativa; seeds; Q6L4H1), maize (Zea mays; tissue not defined; B6UFS4), purple false brome (Brachypodium distachyon; genomic DNA; I1HGT0), sorghum (Sorghum bicolor; genomic DNA; C5YV13), barley (Hordeum vulgare; roots; P93191), rape (B. rapa; anthers; O64431), upland cotton (Gossypium hirsutum; cotton fibers; B6E319) and common wheat (T. aestivum; leaves and seeds; P82900) were aligned using ClustalW2 on EMBL-EBI (B). Scientific names of plants, the tissues where the proteins have been identified and the accession numbers (UniProt) are given in parentheses. Identity scores for each protein are shown in percent. Amino acids identical to those of Api g 6 are highlighted in red and similar residues are shown in yellow.

Figure 2. Alignment of 3-D structure models of Api g 6 (yellow) and Api g 2 (green). Models were generated using rice nsLTP2 (PDA: 1L6H) as template for Api g 6 and Pru p 3 (PDB: 2ALG) as template for Api g 2. Different orientations of the molecules are depicted to show similar alpha-helical secondary structures.

Figure 3. Simulated endolysosomal degradation. Purified natural Api g 6 was incubated with microsomes obtained monocyte-derived dendritic cells of nsLTP-allergic patients and proteolysis was monitored by gel electrophoresis (A). Degradation of intact Api g 6 was evaluated densitometrically and is given in percent (B).

mnfr2033-sup-0002-TableS1.docx25KTable 1.

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