{"doi":"10.1111/tpj.14215","title":"Rapid and reproducible phosphopeptide enrichment by tandem <scp>metal oxide affinity chromatography</scp>: application to boron deficiency induced phosphoproteomics","abstract":"<jats:title>Summary</jats:title><jats:p>Mass spectrometry has been instrumental in enabling the study of molecular signaling on a cellular scale by way of site‐specific quantification of protein post‐translational modifications, in particular phosphorylation. Here we describe an updated tandem metal oxide affinity chromatography (<jats:styled-content style=\"fixed-case\">MOAC)</jats:styled-content> combined phosphoprotein/phosphopeptide enrichment strategy, a scalable phosphoproteomics approach that allows rapid identification of thousands of phosphopeptides in plant materials. We implemented modifications to several steps of the original tandem <jats:styled-content style=\"fixed-case\">MOAC</jats:styled-content> procedure to increase the amount of quantified phosphopeptides and hence site‐specific phosphorylation of proteins in a sample beginning with the less amounts of tissue and a substantially smaller amount of extracted protein. We applied this technology to generate time‐resolved maps of boron signaling in Arabidopsis roots. We show that the successive enrichment of phosphoproteins in a first and phosphopeptide extraction in a second step using our optimized procedure strongly enriched the root phosphoproteome. Our results reveal that boron deficiency affects over 20% of the measured root phosphoproteome and that many phosphorylation sites with known biological function, and an even larger number of previously undescribed sites, are modified during the time course of boron deficiency. We identify transcription factors as key regulators of hormone signaling pathways that modulate gene expression in boron deprived plants. Furthermore, our phosphorylation kinetics data demonstrate that mitogen‐activated protein kinase (<jats:styled-content style=\"fixed-case\">MAPK)</jats:styled-content> cascades mediate polarized transport of boron in Arabidopsis roots. Taken together, we establish and validate a robust approach for proteome‐wide phosphorylation analysis in plant biology research.</jats:p>","journal":"The Plant Journal","year":2019,"id":640588,"datarank":0.4566783656585135,"base_score":3.044522437723423,"endowment":3.044522437723423,"self_citation_contribution":0.4566783656585135,"citation_network_contribution":0.0,"self_endowment_contribution":0.4566783656585135,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":20,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1664928,"name":"Wolfgang Hoehenwarter","orcid":null,"position":1,"is_corresponding":false},{"id":39974,"name":"Yanmei Chen","orcid":"0000-0003-4318-4244","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Rapid and reproducible phosphopeptide enrichment by tandem <scp>metal oxide affinity chromatography</scp>: application to boron deficiency induced phosphoproteomics","abstract":"<jats:title>Summary</jats:title><jats:p>Mass spectrometry has been instrumental in enabling the study of molecular signaling on a cellular scale by way of site‐specific quantification of protein post‐translational modifications, in particular phosphorylation. Here we describe an updated tandem metal oxide affinity chromatography (<jats:styled-content style=\"fixed-case\">MOAC)</jats:styled-content> combined phosphoprotein/phosphopeptide enrichment strategy, a scalable phosphoproteomics approach that allows rapid identification of thousands of phosphopeptides in plant materials. We implemented modifications to several steps of the original tandem <jats:styled-content style=\"fixed-case\">MOAC</jats:styled-content> procedure to increase the amount of quantified phosphopeptides and hence site‐specific phosphorylation of proteins in a sample beginning with the less amounts of tissue and a substantially smaller amount of extracted protein. We applied this technology to generate time‐resolved maps of boron signaling in Arabidopsis roots. We show that the successive enrichment of phosphoproteins in a first and phosphopeptide extraction in a second step using our optimized procedure strongly enriched the root phosphoproteome. Our results reveal that boron deficiency affects over 20% of the measured root phosphoproteome and that many phosphorylation sites with known biological function, and an even larger number of previously undescribed sites, are modified during the time course of boron deficiency. We identify transcription factors as key regulators of hormone signaling pathways that modulate gene expression in boron deprived plants. Furthermore, our phosphorylation kinetics data demonstrate that mitogen‐activated protein kinase (<jats:styled-content style=\"fixed-case\">MAPK)</jats:styled-content> cascades mediate polarized transport of boron in Arabidopsis roots. Taken together, we establish and validate a robust approach for proteome‐wide phosphorylation analysis in plant biology research.</jats:p>","is_dataset_classified":null,"base_score":3.044522437723423,"endowment":3.044522437723423,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"30589143","pmcid":null,"openalex_id":"https://openalex.org/W2908188952","authors":[],"funders":[{"funder_name":"National Natural Science Foundation of China","grant_id":"31270305","title":null},{"funder_name":"National Natural Science Foundation of China","grant_id":"31470345","title":null}],"total_grants":2,"fwci":1.4176,"citation_percentile":0.79133103,"influential_citations":0,"citation_trend":[{"year":2019,"count":1},{"year":2020,"count":4},{"year":2021,"count":6},{"year":2022,"count":4},{"year":2023,"count":1},{"year":2024,"count":2},{"year":2026,"count":2}],"oa_status":"bronze","license":"http://onlinelibrary.wiley.com/termsAndConditions#vor","oa_locations":[{"url":"https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/tpj.14215","host_type":"journal"},{"url":"https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/tpj.14215","host_type":"publisher"},{"url":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Ftpj.14215","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/pdf/10.1111/tpj.14215","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/full-xml/10.1111/tpj.14215","host_type":"publisher"},{"url":"https://doi.org/10.1111/tpj.14215","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/30589143","host_type":"repository"}],"fields_of_study":["Advanced Proteomics Techniques and Applications","Plant Micronutrient Interactions and Effects","Advanced biosensing and bioanalysis techniques"],"mesh_terms":["Boron","Chromatography, Affinity","Oxides","Phosphopeptides","Phosphoproteins","Phosphorylation","Protein Processing, Post-Translational","Signal Transduction","Arabidopsis","Plant Roots","Proteome","Arabidopsis Proteins","Proteomics","Tandem Mass Spectrometry"],"keywords":["Phosphopeptide","Phosphoproteomics","Biology","Phosphorylation","Phosphoprotein","Proteome","Arabidopsis","Proteomics","Protein phosphorylation","Biochemistry","Protein kinase A","Cell biology","Computational biology","Mutant","Gene","Boron","ABA","Moac","Technical Advance"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-07T12:55:14.742557Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}