{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T08:04:29Z","timestamp":1768896269198,"version":"3.49.0"},"reference-count":47,"publisher":"Oxford University Press (OUP)","issue":"18","license":[{"start":{"date-parts":[[2021,5,31]],"date-time":"2021-05-31T00:00:00Z","timestamp":1622419200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/501100000923","name":"Australian Research Council","doi-asserted-by":"publisher","award":["DP150104050"],"award-info":[{"award-number":["DP150104050"]}],"id":[{"id":"10.13039\/501100000923","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000923","name":"Australian Research Council","doi-asserted-by":"publisher","award":["LP150100826"],"award-info":[{"award-number":["LP150100826"]}],"id":[{"id":"10.13039\/501100000923","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Taiwan-ANU Scholarship"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,9,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>C-TERMINALLY ENCODED PEPTIDEs (CEPs) control diverse responses in plants including root development, root system architecture, nitrogen demand signalling, and nutrient allocation that influences yield, and there is evidence that different ligands impart different phenotypic responses. Thus, there is a need for a simple method that identifies bona fide CEP hormone\u2013receptor pairings in vivo and examines whether different CEP family peptides bind the same receptor. We used formaldehyde or photoactivation to cross-link fluorescently tagged group 1 or group 2 CEPs to receptors in semi-purified Medicago truncatula or Arabidopsis thaliana leaf vascular tissues to verify that COMPACT ROOT ARCHITECTURE 2 (CRA2) is the Medicago CEP receptor, and to investigate whether sequence diversity within the CEP family influences receptor binding. Formaldehyde cross-linked the fluorescein isothiocyanate (FITC)-tagged Medicago group 1 CEP (MtCEP1) to wild-type Medicago or Arabidopsis vascular tissue cells, but not to the CEP receptor mutants, cra2 or cepr1. Binding competition showed that unlabelled MtCEP1 displaces FITC\u2013MtCEP1 from CRA2. In contrast, the group 2 CEP, FITC\u2013AtCEP14, bound to vascular tissue independently of CEPR1 or CRA2, and AtCEP14 did not complete with FITC\u2013MtCEP1 to bind CEP receptors. The binding of a photoactivatable FITC\u2013MtCEP1 to the periphery of Medicago vascular cells suggested that CRA2 localizes to the plasma membrane. We separated and visualized a fluorescent 105 kDa protein corresponding to the photo-cross-linked FITC\u2013MtCEP1\u2013CRA2 complex using SDS\u2013PAGE. Mass spectrometry identified CRA2-specific peptides in this protein band. The results indicate that FITC\u2013MtCEP1 binds to CRA2, MtCRA2 and AtCEPR1 are functionally equivalent, and the binding specificities of group 1 and group 2 CEPs are distinct. Using formaldehyde or photoactivated cross-linking of biologically active, fluorescently tagged ligands may find wider utility by identifying CEP\u2013CEP receptor pairings in diverse plants.<\/jats:p>","DOI":"10.1093\/jxb\/erab244","type":"journal-article","created":{"date-parts":[[2021,5,29]],"date-time":"2021-05-29T14:54:37Z","timestamp":1622300077000},"page":"6164-6174","source":"Crossref","is-referenced-by-count":11,"title":["A new method to visualize CEP hormone\u2013CEP receptor interactions in vascular tissue <i>in vivo<\/i>"],"prefix":"10.1093","volume":"72","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9858-757X","authenticated-orcid":false,"given":"Han-Chung","family":"Lee","sequence":"first","affiliation":[{"name":"Division of Plant Sciences, Research School of Biology, College of Science, The Australian National University, Canberra, ACT 2601, 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Australian National University, Canberra, ACT 2601, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4374-1728","authenticated-orcid":false,"given":"Ariel","family":"Ivanovici","sequence":"additional","affiliation":[{"name":"Division of Plant Sciences, Research School of Biology, College of Science, The Australian National University, Canberra, ACT 2601, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4073-8088","authenticated-orcid":false,"given":"John P","family":"Rathjen","sequence":"additional","affiliation":[{"name":"Division of Plant Sciences, Research School of Biology, College of Science, The Australian National University, Canberra, ACT 2601, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9819-1372","authenticated-orcid":false,"given":"Michael 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