Cloning and characterization of 2S albumin, Car i 1, a major allergen in pecan.
Cloning and characterization of 2S albumin, Car i 1, a major allergen in pecan.
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2S白蛋白，CAR I 1的克隆和表征，山核桃的主要过敏原。
DOI：
10.1021/jf104319d
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发表时间：
2011
影响因子：
6.1
通讯作者：
S. Sathe
中科院分区：
文献类型：
--
作者：
G. Sharma;Andre Irsigler;P. Dhanarajan;R. Ayuso;Luda Bardina;H. Sampson;K. Roux;S. Sathe
虽然山核桃与IgE介导的食物过敏有关，但负责的过敏原仍有待鉴定和表征。从山核桃cDNA文库中扩增出2S白蛋白基因。以山核桃过敏患者血清为抗原，用斑点杂交法筛选重组蛋白。通过Edman测序和质谱/质谱（MS/MS）分析来分析亲和纯化的天然蛋白。采用抑制酶联免疫吸附试验（ELISA）测定与核桃的交叉反应性。通过用三个不同患者的血清池探测重叠肽来确定序列表位。三维同源模型的产生，和山核桃表位的位置进行了比较与其他过敏性树坚果同源物上的已知的顺序表位。在通过斑点印迹测试的28名患者中，22名（79%）结合至2S白蛋白，指定为Car i 1。Edman测序和天然2S白蛋白的MS/MS测序证实了重组（r）Car i 1的同一性。山核桃和核桃蛋白提取物均抑制rCar i 1的IgE结合。顺序表位作图表明，弱，中等，和强的反应性对12，7，和5肽，分别。在所有血清库识别的11种肽中，5种肽具有强反应性，并且位于Car i 1的3个离散区域（氨基酸43-57、67-78和106-120）。三维建模显示IgE反应性表位是溶剂可及的，并与其他树坚果具有显著的同源性，为先前观察到的交叉反应性提供了可能的基础。
Although pecans are associated with IgE-mediated food allergies, the allergens responsible remain to be identified and characterized. The 2S albumin gene was amplified from the pecan cDNA library. Dot-blots were used to screen the recombinant protein with pecan allergic patients' serum. The affinity purified native protein was analyzed by Edman sequencing and mass spectrometry/mass spectrometry (MS/MS) analysis. Cross-reactivity with walnut was determined by inhibition enzyme-linked immunosorbent assay (ELISA). Sequential epitopes were determined by probing the overlapping peptides with three different patients' serum pool. The 3-dimensional homology model was generated, and the locations of the pecan epitopes were compared with those of known sequential epitopes on other allergenic tree nut homologues. Of 28 patients tested by dot-blot, 22 (79%) bound to 2S albumin, designated as Car i 1. Edman sequencing and the MS/MS sequencing of native 2S albumin confirmed the identity of recombinant (r) Car i 1. Both pecan and walnut protein extracts inhibited the IgE-binding to rCar i 1. Sequential epitope mapping indicated weak, moderate, and strong reactivity against 12, 7, and 5 peptides, respectively. Of the 11 peptides recognized by all serum pools, 5 peptides were strongly reactive and located in 3 discrete regions of the Car i 1 (amino acids 43-57, 67-78, and 106-120). Three-dimensional modeling revealed IgE-reactive epitopes to be solvent accessible and share significant homology with other tree nuts providing a possible basis for previously observed cross-reactivity.
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影响因子：
2.9
作者：
FLING, SP;GREGERSON, DS
通讯作者：
GREGERSON, DS
影响因子：
14.2
作者：
BURKS, AW;WILLIAMS, LW;HELM, RM
通讯作者：
HELM, RM
影响因子：
14.2
作者：
Sicherer, SH;Furlong, TJ;Sampson, HA
通讯作者：
Sampson, HA
影响因子：
14.2
作者：
Teuber, SS;Dandekar, AM;Sellers, CL
通讯作者：
Sellers, CL
影响因子：
14.2
作者：
Sicherer, SH;Muñoz-Furlong, A;Sampson, HA
通讯作者：
Sampson, HA