Proteomic identification of allergenic proteins in red oak (Quercus rubra) pollen

José Ángel Huerta-Ocampo; Alejandra Valenzuela-Corral; María Del Refugio Robles-Burgueño; Ana María Guzmán-Partida; Miguel Ángel Hernández-Oñate; Luz Vázquez-Moreno; Gandhi F Pavón-Romero; Luis M Terán

doi:10.1016/j.waojou.2020.100111

Proteomic identification of allergenic proteins in red oak (Quercus rubra) pollen

World Allergy Organ J. 2020 Mar 17;13(3):100111. doi: 10.1016/j.waojou.2020.100111. eCollection 2020 Mar.

Authors

José Ángel Huerta-Ocampo¹, Alejandra Valenzuela-Corral², María Del Refugio Robles-Burgueño², Ana María Guzmán-Partida², Miguel Ángel Hernández-Oñate¹, Luz Vázquez-Moreno², Gandhi F Pavón-Romero³, Luis M Terán³

Affiliations

¹ 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.
² 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.
³ Instituto Nacional de Enfermedades Respiratorias "Ismael Cosío Villegas", Calzada Tlalpan No. 4502, Sección XVI, C.P.14080, Ciudad de México, Mexico.

Abstract

Background: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

Keywords: 2-DE, Two-dimensional electrophoresis; AIT, Allergy immunotherapy; BSA, Bovine serum albumin; CHAPS, (3-(3-Cholamidopropyl)dimethylammonio)-1-propanesulfonate); DTT, Dithiothreitol; ED, Emergency department; IEF, Isoelectric focusing; IPG, Immobilized pH gradient; Immunoproteomics; LC, Liquid chromatography; MS, Mass spectrometry; MS/MS, Tandem mass spectrometry; Mass spectrometry; PBS, Phosphate-buffered saline; PMSF, Phenyl methyl sulfonyl fluoride; PVDF, Polyvinylidene difluoride; Pollen allergy; Q-TOF, Quadrupole Time-of-Flight; Red oak; SDS, Sodium dodecyl sulfate; Two-dimensional gel electrophoresis.