Huerta-Ocampo José Ángel, Valenzuela-Corral Alejandra, Robles-Burgueño María Del Refugio, Guzmán-Partida Ana María, Hernández-Oñate Miguel Ángel, Vázquez-Moreno Luz, Pavón-Romero Gandhi F, Terán Luis M
CONACYT-Centro de Investigación en Alimentación y Desarrollo, A.C. Carretera Gustavo Enrique Artizarán Rosas No. 46, Colonia La Victoria, C.P. 83304, Hermosillo, Sonora, Mexico.
Coordinación de Ciencia de los Alimentos, Centro de Investigación en Alimentación y Desarrollo, A.C. Carretera Gustavo Enrique Artizarán Rosas No. 46, Colonia La Victoria, C.P. 83304, Hermosillo, Sonora, Mexico.
World Allergy Organ J. 2020 Mar 17;13(3):100111. doi: 10.1016/j.waojou.2020.100111. eCollection 2020 Mar.
Red oak pollen is an important cause of allergic respiratory disease and it is widely distributed in North America and central Europe. To date, however, red oak pollen allergens have not been identified. Here, we describe the allergenic protein profile from red oak pollen.
Total proteins were extracted from red oak pollen using a modified phenolic extraction method, and, subsequently, proteins were separated by two-dimensional gel electrophoresis (2DE) for both total protein stain (Coomassie Blue) and immunoblotting. A pool of 8 sera from red oak sensitive patients was used to analyze blotted proteins. Protein spots were analyzed by Mass Spectrometry.
Electrophoretic pattern of total soluble proteins showed higher intensity bands in the regions of 26-40 and 47-52 kDa. Two dimensional immunoblots using pool sera from patients revealed four allergenic proteins spots with molecular masses in the range from 50 to 55 kDa. Mass spectrometry analysis identified 8 proteins including Enolase 1 and Enolase 1 chloroplastic, Xylose isomerase (X1 isoform), mitochondrial Aldehyde dehydrogenase, UTP-Glusose-1-phosphate uridylyltransferase, Betaxylosidase/alpha-l-arabinofuranosidase and alpha- and beta subunits of ATP synthase.
This study has identified for first time 8 IgE binding proteins from red oak pollen. These findings will pave the way towards the development of new diagnostic and therapeutic modalities for red oak allergy.
红橡木花粉是过敏性呼吸道疾病的一个重要诱因，在北美和中欧广泛分布。然而，迄今为止，红橡木花粉过敏原尚未得到鉴定。在此，我们描述了红橡木花粉的致敏蛋白图谱。
采用改良的酚提取法从红橡木花粉中提取总蛋白，随后，通过二维凝胶电泳（2DE）对总蛋白进行染色（考马斯亮蓝）和免疫印迹分析。使用来自对红橡木敏感患者的8份血清混合样本分析印迹蛋白。通过质谱分析蛋白斑点。
总可溶性蛋白的电泳图谱显示，在26 - 40 kDa和47 - 52 kDa区域有强度较高的条带。使用患者血清混合样本进行的二维免疫印迹显示有4个致敏蛋白斑点，分子量在50至55 kDa范围内。质谱分析鉴定出8种蛋白质，包括烯醇化酶1和叶绿体烯醇化酶1、木糖异构酶（X1亚型）、线粒体醛脱氢酶、UTP - 葡萄糖 - 1 - 磷酸尿苷转移酶、β - 木糖苷酶/α - L - 阿拉伯呋喃糖苷酶以及ATP合酶的α和β亚基。
本研究首次鉴定出红橡木花粉中的8种IgE结合蛋白。这些发现将为红橡木过敏新诊断和治疗方法的开发铺平道路。
