[Analysis of the electrophoretic profiles of the protein extracts of the species Manihot esculenta (Crantz, 1975), Persea Americana (Mill., 1768) and Actinidia delicious (A. Chevalier., 1984), and their association with the latex-fruit syndrome for searching for potential allergens].

Alfredo Lagares, Juliana Jaspe, Daibeth Domínguez, Angela Muñoz, Eduardo Egea
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Abstract

Objective: Determine the electrophoretic profiles of the extracts of Manihot esculenta, Actinidia Deliciosa and Persea Americana and their possible relationship with Latex-Fruit Syndrome.

Methods: Protein extracts of M. esculenta, P. Americana and A. Deliciosa were prepared through the processes of maceration and solvent extraction from plant samples. In the case of the avocado, a prior extraction by soxhlet was carried out to eliminate the fat. The extracts were vacuum filtered, dialyzed and finally lyophilized. Separation of proteins based on molecular weight was performed by SDS PAGE electrophoresis. The electrophoretic profiles obtained were compared with the allergenic proteins previously identified in the latex extract, in order to determine a possible relationship with Latex-Fruit Syndrome, depending on the molecular weight.

Results: The extracts of M. esculenta and P. Americana showed a wide range of protein fractions with molecular weights varying from 10 to 250 KD, finding that the region with the highest concentration of bands was between 20 and 89 KD, (60 and 65%), respectively. A 20-band profile was obtained for the M. esculenta extract (Figure 1), with seven bands sharing similar weights with the latex allergens (Hev b 1, Hev b 2, Hev b3, Hev b 4, Hev b 5, Hev b 6.03, Hev b 8 and Hev b 10) (3-5). For the P. Americana extract, 20 bands were also observed (Figure 2), seven of which presented approximate weights to the Latex allergens (Hev b 1, Hev b 2 Hev b 4 Hev b 6.01 Hev b 6.03 Hev b 8 , Hev b 10 Hev b 11 Hev b 14). The Kiwi extract showed two bands of 19.1 and 22.9 KD, with weights close to latex proteins (figure 3), (Hev b 3 and Hev b 6.01), and allergens (Act d 2 and Act d 6), reported in the literature for this fruit.

Conclusions: When analyzing the relationship between the separated protein fractions and the latex allergens described in the literature, a possible association of 35% was found for the extracts of M. esculenta and P. Americana, and 10% for A. Delicious, with great relevance being the association found with the allergens Hev b 4, Hev b 2, Hev 8 and Hev b 11, which are involved in Latex-Fruit Syndrome. The electrophoretic profiles of the prepared extracts were determined and compared with the Latex allergens. This information generates a contribution for the development of new research and advances in the standardization of these extracts on a large scale and for their future use in diagnostic tests.

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[分析 Manihot esculenta(Crantz,1975 年)、Persea Americana(Mill.,1768 年)和 Actinidia delicious(A. Chevalier.,1984 年)物种蛋白质提取物的电泳图谱,以及它们与乳胶果综合症的关联,以寻找潜在的过敏原]。
目的确定 Manihot esculenta、Actinidia Deliciosa 和 Persea Americana 提取物的电泳图谱及其与乳汁果实综合征的可能关系:方法:通过浸渍和溶剂萃取植物样本的方法,制备了 Manihot esculenta、P. Americana 和 A. Deliciosa 的蛋白质提取物。在牛油果中,先用索氏提取法去除脂肪。提取物经过真空过滤、透析,最后冻干。通过 SDS PAGE 电泳对蛋白质进行分子量分离。将获得的电泳图谱与之前在乳胶提取物中发现的致敏蛋白质进行比较,以确定与乳胶水果综合症之间可能存在的关系(取决于分子量):M. esculenta 和 P. Americana 的提取物显示出分子量从 10 到 250 KD 不等的多种蛋白质组分,发现条带浓度最高的区域分别在 20 到 89 KD 之间(60% 和 65%)。在 M. esculenta 提取物中得到了 20 条带图谱(图 1),其中 7 条带与乳胶过敏原(Hev b 1、Hev b 2、Hev b 3、Hev b 4、Hev b 5、Hev b 6.03、Hev b 8 和 Hev b 10)具有相似的重量(3-5)。在美洲豹提取物中也观察到 20 个条带(图 2),其中 7 个条带与乳胶过敏原近似(Hev b 1、Hev b 2、Hev b 4、Hev b 6.01、Hev b 6.03、Hev b 8、Hev b 10、Hev b 11、Hev b 14)。猕猴桃提取物中出现了两条分别为 19.1 和 22.9 KD 的条带,其重量与文献中报道的该水果的乳胶蛋白(图 3)(Hev b 3 和 Hev b 6.01)和过敏原(Act d 2 和 Act d 6)相近:在分析分离的蛋白质组分与文献中描述的乳胶过敏原之间的关系时,发现 M. esculenta 和 P. Americana 的提取物可能与 35% 的过敏原有关,A. Delicious 的提取物可能与 10% 的过敏原有关,其中与乳胶水果综合症过敏原 Hev b 4、Hev b 2、Hev 8 和 Hev b 11 的关系最为密切。对制备的提取物的电泳图谱进行了测定,并与乳胶过敏原进行了比较。这些信息有助于开展新的研究,推动这些提取物的大规模标准化,并促进它们在未来诊断测试中的应用。
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