Mutational analysis of the IgE-binding epitopes of P34/Gly m Bd 30K
Mutational analysis of the IgE-binding epitopes of P34/Gly m Bd 30K
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DOI：
10.1016/s0091-6749(00)90091-5
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发表时间：
2000-02-01
影响因子：
14.2
通讯作者：
Burks, AW
中科院分区：
文献类型：
--
作者：
Helm, RM;Cockrell, G;Burks, AW
背景：花生和大豆是两种食物，已被证明是负责许多特应性疾病。由于其营养价值，大豆蛋白现在越来越多地用于许多食品中。以往的研究已经证明大豆提取物中存在多种过敏原，包括大豆球蛋白、β-伴大豆球蛋白和P34/Gly m Bd 30 K蛋白。目的：我们的总体目标是鉴定大豆特异性过敏原，以便开始了解豆类蛋白的分子和免疫化学特征。本研究的具体目的是鉴定P34/Gly m艾德30 K的5个不同免疫显性表位中IgE结合所必需的必需氨基酸残基。用6名临床敏感的大豆过敏个体的血清IgE来鉴定P34/Gly m艾德30 K在天然肽和单氨基酸取代肽中的含量，使用SPOTS肽合成技术测定Gly m 30 K所需的关键氨基酸，IgE结合。来自个体的血清IgE的IgE结合和表位识别的强度变化很大。结论：Gly m艾德30 K的5个免疫优势表位经丙氨酸单位点置换后，可使6例大豆敏感患者血清中表位6和16的IgE结合能力降低或消失。
Background: Peanuts and soybeans are 2 foods that have been shown to be responsible for many atopic disorders. Because of their nutritional benefit, soybean proteins are now being used increasingly in a number of food products. Previous studies have documented multiple allergens in soybean extracts, including glycinin, beta-conglycinin, and the P34/Gly m Bd 30K protein.Objective: Our overall goal was to identify soybean-specific allergens to begin to understand molecular and immunochemical characteristics of legume proteins. The specific aim of the current investigation was to identify the essential amino acid residues necessary for IgE binding in the 5 distinct immunodominant epitopes of P34/Gly m Ed 30K,Methods: Serum IgE from 6 clinically sensitive soybean-allergic individuals was used to identify P34/Gly m Ed 30K in the native and single amino acid substituted peptides with use of the SPOTS peptide synthesis technique to determine critical amino acids required for IgE binding.Results: The intensity of IgE binding and epitope recognition by serum IgE from the individuals varied substantially. With use of serum from 6 clinically soybean-sensitive individuals, 2 of the 5 immunodominant epitopes could be mutagenized to non-IgE binding peptides,Conclusions: Single-site amino acid substitution of the 5 immunodominant epitopes of Gly m Ed 30K with alanine revealed that IgE binding could be reduced or eliminated in epitopes 6 and 16 in the serum obtained from 6 soybean-sensitive patients.