Purification and Characterization of Ara h1 and Ara h3 from Four Peanut Market Types Revealed Higher Order Oligomeric Structures

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• 作者：Eriyanto Yusnawan ; Christopher P. Marquis ; N. Alice Lee
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2012
• 出版时间：October 17, 2012
• 年：2012
• 卷：60
• 期：41
• 页码：10352-10358
• 全文大小：288K
• 年卷期：v.60,no.41(October 17, 2012)
• ISSN：1520-5118

文摘

This study aimed to purify and characterize the peanut allergens Ara h1 and Ara h3 from four cultivars that represent the four major market types to provide better understanding of the molecular organization of oligomers in different market types. The chromatographic profiles of Ara h1 and Ara h3 from the four cultivars obtained from anion exchange chromatography were similar. However, they differed in the distribution of trimeric and hexameric structures of Ara h3 isolated by size exclusion chromatography. The Menzies (Runner market type) and Walter (Spanish market type) cultivars, wherein Ara h3 proteins consist of two acidic subunits, exhibited trimeric and hexameric conformations proportionally. However, the Middleton (Virginia market type) and Kelinci (Valencia market type) cultivars, wherein Ara h3 proteins consist of three acidic subunits, showed predominantly a hexameric structure. The oligomeric structures of the purified Ara h1 demonstrated strong IgE binding properties, whereas the allergenic property of the oligomeric Ara h3 could not be performed due to lack of availability of specific IgE. In addition, the polyclonal antibodies raised against the purified Ara h1 and Ara h3 showed highly specific binding to their respective antigens.

Keywords:

anion exchange chromatography; size exclusion chromatography; peanut allergens; Ara h1; Ara h3; 2D electrophoresis; mass spectrometry; human IgE antibodies; rabbit IgG antibodies