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中国精品科技期刊2020
张爱琳,赵慧娟,迟越,等. 腰果过敏蛋白Ana o 2与Ana o 3结构分析及致敏性预测[J]. 华体会体育,2022,43(5):34−40. doi: 10.13386/j.issn1002-0306.2021010189.
引用本文: 张爱琳,赵慧娟,迟越,等. 腰果过敏蛋白Ana o 2与Ana o 3结构分析及致敏性预测[J]. 华体会体育,2022,43(5):34−40. doi: 10.13386/j.issn1002-0306.2021010189.
ZHANG Ailin, ZHAO Huijuan, CHI Yue, et al. Structural Analysis and Sensitization Prediction of the Allergic Proteins Ana o 2 and Ana o 3 in Cashew Nuts[J]. Science and Technology of Food Industry, 2022, 43(5): 34−40. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021010189.
Citation: ZHANG Ailin, ZHAO Huijuan, CHI Yue, et al. Structural Analysis and Sensitization Prediction of the Allergic Proteins Ana o 2 and Ana o 3 in Cashew Nuts[J]. Science and Technology of Food Industry, 2022, 43(5): 34−40. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021010189.

腰果过敏蛋白Ana o 2与Ana o 3结构分析及致敏性预测

Structural Analysis and Sensitization Prediction of the Allergic Proteins Ana o 2 and Ana o 3 in Cashew Nuts

  • 摘要: 为了解腰果主要过敏蛋白的分子结构及过敏性特征,通过有机溶剂脱脂、盐析、透析、葡聚糖凝胶Sephadex G-150分离纯化得到腰果主要过敏蛋白,采用紫外分光光度计、BCA定量试剂盒、十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)、蛋白质免疫印迹(Western-Blotting)等手段检测腰果过敏蛋白及评价其免疫原性;应用NCBI数据库与蛋白质谱测序分析获得腰果过敏蛋白氨基酸序列、DNA序列,结合生物信息软件DNAstar与圆二色谱(CD)谱图预测腰果过敏蛋白结构。结果表明:Ana o 2与Ana o 3为腰果主要过敏蛋白,分子量分别为33和17 kD左右,根据圆二色谱检测结果与DNAstar中的Protean程序预测分析可知,Ana o 2中α-螺旋、β-折叠区段和转角区域均存在,是致敏性较强的一种亲水性蛋白,β-转角和无规则卷曲在Ana o 2氨基酸序列中出现率总和为54%,所在区域为优势抗原表位区。Ana o 3仅有少量的 β-折叠区段和转角区域,出现率总和为12%,Ana o 3的二级结构主要为 α-螺旋结构和无规则卷曲结构,不适宜作为抗原的表位,形成过敏性表位相对较弱。本研究结果为进一步明确天然腰果主要过敏原结构并对探索腰果致敏机理提供参考。

     

    Abstract: In order to definite the molecular structure and allergic characteristics of cashew nuts, the main allergic proteins were isolated and purified by organic solvent defatting, salting out, dialysis and Sephadex G-150 gel. The allergenic proteins were definitely by UV, BCA kit, SDS-PAGE and Western-Blotting. The amino acid sequence and the DNA sequence of cashew allergic protein were obtained by using the NCBI database and protein mass spectrum sequencing. The structure of cashew allergic protein was predicted by DNAstar bio-information software and circular dichroism (CD). The results showed that the Ana o 2 and Ana o 3 were the main cashew allergenic proteins with molecular weight of 33 and 17 kD. The results of CD and Protean program in DNAstar predicted that there were α-helical, β-folded regions and corner regions which was a hydrophilic protein with strong sensitization because of the β-corners and random curls were found in 54% in Ana o 2. The secondary structure had only a small amount of β-folded region and corner region, accounted for 12%, but had mainly α-helix and random coil structure. It was also hydrophilic allergic protein, and the formation of allergenic epitopes was relatively weak. This study would provide reference for further clarifying the structure of the main allergens of natural cashew nuts and exploring the sensitization mechanism of cashew nuts.

     

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