Isoform identification and characterization of Art v 3, the lipid-transfer protein of mugwort pollen

被引：42

作者：

Gadermaier, Gabriele ^{[1
]}

Harrer, Andrea ^{[2
]}

Girbl, Tamara ^{[1
]}

Palazzo, Paola ^{[3
]}

Himly, Martin ^{[2
]}

Vogel, Lothar ^{[4
]}

Briza, Peter ^{[2
]}

Mari, Adriano ^{[3
]}

Ferreira, Fatima ^{[1
,2
]}

机构：

[1] Salzburg Univ, Dept Mol Biol, Christian Doppler Lab Allergy Diag & Therapy, A-5020 Salzburg, Austria

[2] Salzburg Univ, Dept Mol Biol, Div Allergy & Immunol, A-5020 Salzburg, Austria

[3] IRCCS, IDI, Ctr Clin & Expt Allergol, I-00167 Rome, Italy

[4] Paul Ehrlich Inst, Div Allergol, D-63225 Langen, Germany

来源：

MOLECULAR IMMUNOLOGY | 2009年 / 46卷 / 10期

关键词：

Art v 3; Cross-reactivity; Mugwort pollen allergy; Isoforms; Lipid-transfer protein; IGE-BINDING EPITOPES; CROSS-REACTIVITY; PLANT FOODS; CLINICAL EXPRESSION; RESPIRATORY ALLERGY; ARTEMISIA POLLEN; ROSACEAE FRUITS; PEACH PRU-P-3; DIGESTION; FAMILY;

D O I：

10.1016/j.molimm.2009.03.021

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Art v 3, the lipid-transfer protein (LTP) of Artemisia vulgaris pollen is a relevant allergen showing frequent cross-reactivity with homologues in other plants. Here we report the identification of four full-length Art v 3 sequences obtained by cDNA cloning using mass spectrometry-based sequencing. Two isoforms, Art v 3.0201 and Art v 3.0301 were expressed as soluble proteins in Escherichia coli Rosetta-gami B(DE3) pLysS using different expression systems. Purified natural and recombinant Art v 3 demonstrated similar secondary structures in circular dichroism analysis. All preparations showed high thermal stability but low resistance to gastric digestion with pepsin. Patient-specific IgE reactivity patterns to natural or recombinant isoallergens were observed among Art v 3-sensitized subjects. Using Immuno Solid-phase Allergen Chip (ISAC) assays, frequent cross-reactivity of Art v 3 with LTPs from peach and hazelnut was shown. The biological activity of both isoforms was comparable to the natural allergen in basophil release assays. The newly identified sequences provide the basis for recombinant mugwort LTP production enabling batch-to-batch reproducibility and thus ensuring high-quality products for diagnosis and therapy. (C) 2009 Elsevier Ltd. All rights reserved.

引用

页码：1919 / 1924

页数：6

共 37 条

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