{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T08:26:30Z","timestamp":1776155190614,"version":"3.50.1"},"reference-count":17,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2021,9,25]],"date-time":"2021-09-25T00:00:00Z","timestamp":1632528000000},"content-version":"am","delay-in-days":365,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#am"},{"start":{"date-parts":[[2020,9,25]],"date-time":"2020-09-25T00:00:00Z","timestamp":1600992000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["R01GM121445"],"award-info":[{"award-number":["R01GM121445"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003627","name":"Rural Development Administration","doi-asserted-by":"publisher","award":["PJ01325403"],"award-info":[{"award-number":["PJ01325403"]}],"id":[{"id":"10.13039\/501100003627","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["currentprotocols.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["CP Plant Biology"],"published-print":{"date-parts":[[2020,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Protein S\u2010acylation, predominately in the form of palmitoylation, is a reversible lipid post\u2010translational modification on cysteines that plays important roles in protein localization, trafficking, activity, and complex assembly. The functions and regulatory mechanisms of S\u2010acylation have been extensively studied in mammals owing to remarkable development of high\u2010resolution proteomics and the discovery of the S\u2010acylation\u2010related enzymes. However, the advancement of S\u2010acylation studies in plants lags behind that in mammals, mainly due to the lack of knowledge about proteins responsible for this process, such as protein acyltransferases and their substrates. In this article, a set of systematic protocols to study global S\u2010acylation in <jats:italic>Arabidopsis<\/jats:italic> seedlings is described. The procedures are presented in detail, including preparation of <jats:italic>Arabidopsis<\/jats:italic> seedlings, enrichment of plasma membrane (PM) proteins, ensuing enrichment of S\u2010acylated proteins\/peptides based on the acyl\u2010biotin exchange method, and large\u2010scale identification of S\u2010acylated proteins\/peptides via mass spectrometry. This approach enables researchers to study S\u2010acylation of PM proteins in plants in a systematic and straightforward way. \u00a9 2020 Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Preparation of <jats:italic>Arabidopsis<\/jats:italic> seedling materials<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: Isolation and enrichment of plasma membrane proteins<\/jats:p><jats:p><jats:bold>Support Protocol 1<\/jats:bold>: Determination of protein concentration using BCA assay<\/jats:p><jats:p><jats:bold>Basic Protocol 3<\/jats:bold>: Enrichment of S\u2010acylated proteins by acyl\u2010biotin exchange method<\/jats:p><jats:p><jats:bold>Support Protocol 2<\/jats:bold>: Protein precipitation by methanol\/chloroform method<\/jats:p><jats:p><jats:bold>Basic Protocol 4<\/jats:bold>: Trypsin digestion and proteomic analysis<\/jats:p><jats:p><jats:bold>Alternate Protocol<\/jats:bold>: Pre\u2010resin digestion and peptide\u2010level enrichment<\/jats:p>","DOI":"10.1002\/cppb.20119","type":"journal-article","created":{"date-parts":[[2020,9,25]],"date-time":"2020-09-25T16:31:25Z","timestamp":1601051485000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Selective Enrichment Coupled with Proteomics to Identify S\u2010Acylated Plasma Membrane Proteins in <i>Arabidopsis<\/i>"],"prefix":"10.1002","volume":"5","author":[{"given":"Lijuan","family":"Zhou","sequence":"first","affiliation":[{"name":"Division of Plant Sciences, C.S. Bond Life Science Center University of Missouri  Columbia Missouri"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mowei","family":"Zhou","sequence":"additional","affiliation":[{"name":"Environmental Molecular Sciences Laboratory Pacific Northwest National Laboratory  Richland Washington"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marina A.","family":"Gritsenko","sequence":"additional","affiliation":[{"name":"Biological Science Division Pacific Northwest National Laboratory  Richland Washington"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gary","family":"Stacey","sequence":"additional","affiliation":[{"name":"Division of Plant Sciences, C.S. Bond Life Science Center University of Missouri  Columbia Missouri"},{"name":"Division of Biochemistry, C.S. Bond Life Science Center University of Missouri  Columbia Missouri"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,9,25]]},"reference":[{"key":"e_1_2_11_2_1","first-page":"720482","article-title":"S\u2010acylation of plant immune receptors mediates immune signaling in plasma membrane nanodomains","author":"Chen D.","year":"2019","journal-title":"bioRxiv"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-6813-8_13"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-04580-1"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.2144\/04362RR02"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1105\/tpc.17.00044"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.201000197"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1194\/jlr.D011106"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/nph.12077"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.07-9199com"},{"key":"e_1_2_11_11_1","first-page":"346","article-title":"Progress toward understanding protein S\u2010acylation: Prospective in plants","volume":"8","author":"Li Y.","year":"2017","journal-title":"Frontiers in Plant Science"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pbi.2017.07.007"},{"key":"e_1_2_11_13_1","first-page":"249","article-title":"Arabidopsis protein S\u2010acyl transferase 4 mediates root hair growth","volume":"90","author":"Wan Z. 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