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

• Published in: The World Allergy Organization journal Vol. 13; no. 3; p. 100111
• Main Authors: 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.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-03-2020; Elsevier BV; World Allergy Organization; Elsevier
• Subjects: Allergens; Allergies; Asthma; Immunoproteomics; Mass spectrometry; Membranes; Pollen; Pollen allergy; Proteins; Red oak; Two-dimensional gel electrophoresis; Mexico; California; United States > US; Europe; PBS; IPG; Red oak; SDS; Q-TOF; DTT; Immunoproteomics; MS; 2-DE; IEF; AIT; MS/MS; PVDF; CHAPS; BSA; Pollen allergy; PMSF; Two-dimensional gel electrophoresis; LC; Mass spectrometry; ED; SDS, Sodium dodecyl sulfate; CHAPS, (3-(3-Cholamidopropyl)dimethylammonio)-1-propanesulfonate; IPG, Immobilized pH gradient; IEF, Isoelectric focusing; Q-TOF, Quadrupole Time-of-Flight; BSA, Bovine serum albumin; PVDF, Polyvinylidene difluoride; ED, Emergency department; PMSF, Phenyl methyl sulfonyl fluoride; LC, Liquid chromatography; AIT, Allergy immunotherapy; DTT, Dithiothreitol; 2-DE, Two-dimensional electrophoresis; MS, Mass spectrometry; MS/MS, Tandem mass spectrometry; PBS, Phosphate-buffered saline
• ISSN: 1939-4551; 1939-4551
• DOI: 10.1016/j.waojou.2020.100111

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.