聚半乳糖醛酸酶（果胶酶），一种新的油菜籽过敏原。

Polygalacturonase (pectinase), a new oilseed rape allergen.

作者信息

Chardin H, Mayer C, Sénéchal H, Poncet P, Clément G, Wal J M, Desvaux F X, Peltre G

机构信息

Institut Pasteur, Paris, France; Université René Descartes, Paris, France.

出版信息

Allergy. 2003 May;58(5):407-11. doi: 10.1034/j.1398-9995.2003.00094.x.

DOI:10.1034/j.1398-9995.2003.00094.x

PMID:12752327

Abstract

BACKGROUND

Type I hypersensitivity to rapeseed pollen allergens was described as the result of a cross-sensitization with various pollens that could constitute an aggravating factor in birch or grass pollen allergies. Recently, a few rapeseed pollen allergens were described. The aim of the present work was to identify new rapeseed pollen allergens by using two-dimensional gel analysis, microsequencing, and mass spectrometry.

METHODS

Water extractable proteins from oilseed rape pollen or stamen were separated by two-dimensional gel electrophoresis. The proteins were then electroblotted onto a nitrocellulose (NC) sheet. The NC sheets were successively incubated with (1) individual human sera pre-selected for their immunoglobulin E (IgE) reactivity to rapeseed pollen proteins, (2) alkaline phosphatase (AP)-conjugated goat anti-human IgE and (3) AP substrate. The allergens localized by this method were then identified by microsequencing and MALDI-TOF mass spectrometry analysis.

RESULTS

Of the 18 sera studied, five recognized a wide multispot zone with a molecular mass around 43 kD and pIs between 6.5 and 8.5. The results obtained with two representative sera are shown. From this zone, two isoforms of the polygalacturonase enzyme were identified by microsequencing. Confirmation was obtained through MALDI-TOF mass spectrometry analysis.

CONCLUSION

The present results allow the identification of a new rapeseed allergen that can be the main allergen for some patients.

摘要

背景

对油菜花粉过敏原的I型超敏反应被描述为与各种花粉交叉致敏的结果，这些花粉可能是桦树或草花粉过敏的加重因素。最近，有几种油菜花粉过敏原被描述。本研究的目的是通过二维凝胶分析、微量测序和质谱法鉴定新的油菜花粉过敏原。

方法

用二维凝胶电泳分离油菜花粉或雄蕊中的水溶性蛋白质。然后将蛋白质电转移到硝酸纤维素（NC）膜上。NC膜依次与（1）预先选择的对油菜花粉蛋白具有免疫球蛋白E（IgE）反应性的个体人血清、（2）碱性磷酸酶（AP）偶联的山羊抗人IgE和（3）AP底物孵育。通过微量测序和基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱分析鉴定通过该方法定位的过敏原。