PMID- 30856452 OWN - NLM STAT- MEDLINE DCOM- 20191030 LR - 20191030 IS - 1873-6750 (Electronic) IS - 0160-4120 (Linking) VI - 126 DP - 2019 May TI - In-depth quantitative profiling of post-translational modifications of Timothy grass pollen allergome in relation to environmental oxidative stress. PG - 644-658 LID - S0160-4120(18)32266-9 [pii] LID - 10.1016/j.envint.2019.03.001 [doi] AB - An association between pollution (e.g., from traffic emissions) and the increased prevalence of respiratory allergies has been observed. Field-realistic exposure studies provide the most relevant assessment of the effects of the intensity and diversity of urban and industrial contamination on pollen structure and allergenicity. The significance of in-depth post-translational modification (PTM) studies of pollen proteomes, when compared with studies on other aspects of pollution and altered pollen allergenicity, has not yet been determined; hence, little progress has been made within this field. We undertook a comprehensive comparative analysis of multiple polluted and environmentally preserved Phleum pratense (Timothy grass) pollen samples using scanning electron microscopy, in-depth PTM profiling, determination of organic and inorganic pollutants, analysis of the release of sub-pollen particles and phenols/proteins, and analysis of proteome expression using high resolution tandem mass spectrometry. In addition, we used quantitative enzyme-linked immunosorbent assays (ELISA) and immunoglobulin E (IgE) immunoblotting. An increased phenolic content and release of sub-pollen particles was found in pollen samples from the polluted area, including a significantly higher content of mercury, cadmium, and manganese, with irregular long spines on pollen grain surface structures. Antioxidative defense-related enzymes were significantly upregulated and seven oxidative PTMs were significantly increased (methionine, histidine, lysine, and proline oxidation; tyrosine glycosylation, lysine 4-hydroxy-2-nonenal adduct, and lysine carbamylation) in pollen exposed to the chemical plant and road traffic pollution sources. Oxidative modifications affected several Timothy pollen allergens; Phl p 6, in particular, exhibited several different oxidative modifications. The expression of Phl p 6, 12, and 13 allergens were downregulated in polluted pollen, and IgE binding to pollen extract was substantially lower in the 18 patients studied, as measured by quantitative ELISA. Quantitative, unrestricted, and detailed PTM searches using an enrichment-free approach pointed to modification of Timothy pollen allergens and suggested that heavy metals are primarily responsible for oxidative stress effects observed in pollen proteins. CI - Copyright (c) 2019 The Authors. Published by Elsevier Ltd.. All rights reserved. FAU - Smiljanic, Katarina AU - Smiljanic K AD - University of Belgrade-Faculty of Chemistry, Centre of Excellence for Molecular Food Sciences and Department of Biochemistry, Belgrade, Serbia. Electronic address: katarinas@chem.bg.ac.rs. FAU - Prodic, Ivana AU - Prodic I AD - Innovation Center Ltd, University of Belgrade-Faculty of Chemistry, Belgrade, Serbia. FAU - Apostolovic, Danijela AU - Apostolovic D AD - Karolinska Institute, Department of Medicine, Solna, Stockholm, Sweden. FAU - Cvetkovic, Anka AU - Cvetkovic A AD - Institute of Public Health of Belgrade, Belgrade, Serbia. FAU - Veljovic, Djordje AU - Veljovic D AD - University of Belgrade-Faculty of Technology and Metallurgy, Belgrade, Serbia. FAU - Mutic, Jelena AU - Mutic J AD - University of Belgrade-Faculty of Chemistry, Centre of Excellence for Molecular Food Sciences and Department of Biochemistry, Belgrade, Serbia; Ghent University Global Campus, Incheon, South Korea. FAU - van Hage, Marianne AU - van Hage M AD - Karolinska Institute, Department of Medicine, Solna, Stockholm, Sweden. FAU - Burazer, Lidija AU - Burazer L AD - Institute of Immunology, Virology and Sera Production, Torlak Institut, Belgrade, Serbia. FAU - Cirkovic Velickovic, Tanja AU - Cirkovic Velickovic T AD - University of Belgrade-Faculty of Chemistry, Centre of Excellence for Molecular Food Sciences and Department of Biochemistry, Belgrade, Serbia; Ghent University Global Campus, Incheon, South Korea; Ghent University, Faculty of Bioscience Engineering, Ghent, Belgium; Serbian Academy of Sciences and Arts, Belgrade, Serbia. Electronic address: tcirkov@chem.bg.ac.rs. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190308 PL - Netherlands TA - Environ Int JT - Environment international JID - 7807270 RN - 0 (Allergens) RN - 0 (Environmental Pollutants) RN - 0 (Plant Proteins) RN - 0 (Proteome) SB - IM MH - Adult MH - Allergens/*analysis MH - Environmental Pollutants/*analysis MH - Female MH - Humans MH - Male MH - Middle Aged MH - Oxidative Stress MH - Phleum/*physiology MH - Plant Proteins/*metabolism MH - Pollen/*chemistry MH - *Protein Processing, Post-Translational MH - Proteome/*metabolism MH - Serbia MH - Traffic-Related Pollution/analysis OTO - NOTNLM OT - Air-related traffic pollution OT - Allergy OT - Heavy metal pollution OT - Label free relative quantification OT - Oxidative post-translational modification OT - Timothy grass pollen EDAT- 2019/03/12 06:00 MHDA- 2019/10/31 06:00 CRDT- 2019/03/12 06:00 PHST- 2018/10/04 00:00 [received] PHST- 2019/02/02 00:00 [revised] PHST- 2019/03/01 00:00 [accepted] PHST- 2019/03/12 06:00 [pubmed] PHST- 2019/10/31 06:00 [medline] PHST- 2019/03/12 06:00 [entrez] AID - S0160-4120(18)32266-9 [pii] AID - 10.1016/j.envint.2019.03.001 [doi] PST - ppublish SO - Environ Int. 2019 May;126:644-658. doi: 10.1016/j.envint.2019.03.001. Epub 2019 Mar 8.