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We identified linear B-cell epitopes of Pin y 2 using five bioinformatics tools, followed by indirect ELISA with rabbit anti-Pin y 2 IgG and sera from pine nut-allergic patients. Four IgE epitopes and three IgG epitopes were validated. All epitopes are surface-located and predominantly in negatively charged electrostatic regions. Alanine scanning revealed that leucine (L97, L98), glutamic acid (E419, E434, E436), and glutamine (Q442) are key residues, with their alanine substitution causing a reduction of more than 50% in IgE\/IgG binding OD values. Comparative epitope mapping with other tree nut vicilin allergens showed the positional conservation but sequence divergence of the Pin y 2 epitope. These findings reveal the epitope map and epitope key residues of Pin y 2, which provides rational targets for hypoallergenic design and diagnostics.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c06151","type":"journal-article","created":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T01:50:42Z","timestamp":1784771442000},"page":"23816-23826","source":"Crossref","is-referenced-by-count":0,"title":["Identification\nand Key Amino Acid Analysis of Epitopes\nof Pin y 2, a Novel Allergen from\n                    <i>Pinus yunnanensis<\/i>"],"prefix":"10.1021","volume":"74","author":[{"given":"Yuhao","family":"Huang","sequence":"first","affiliation":[{"name":"Ocean University of China , , Sansha Road 1299 , ,","place":["Qingdao, PR China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunying","family":"Sun","sequence":"additional","affiliation":[{"name":"Ocean University of China , , Sansha Road 1299 , ,","place":["Qingdao, PR China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5059-5354","authenticated-orcid":true,"given":"Lirui","family":"Sun","sequence":"additional","affiliation":[{"name":"Qingdao University , , Ningxia Road 308 , ,","place":["Qingdao, PR China, 266071"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiang","family":"Gao","sequence":"additional","affiliation":[{"name":"The Affiliated Hospital of Qingdao University , , Jiangsu Road 16 , ,","place":["Qingdao, PR China, 266003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1016-3976","authenticated-orcid":true,"given":"Hao","family":"Wang","sequence":"additional","affiliation":[{"name":"Ocean University of China , , Sansha Road 1299 , ,","place":["Qingdao, PR China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9881-3565","authenticated-orcid":true,"given":"Ziye","family":"Zhang","sequence":"additional","affiliation":[{"name":"Ocean University of China , , Sansha Road 1299 , ,","place":["Qingdao, PR China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Lin","sequence":"additional","affiliation":[{"name":"Ocean University of China , , Sansha Road 1299 , ,","place":["Qingdao, PR China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7866-681X","authenticated-orcid":true,"given":"Zhenxing","family":"Li","sequence":"additional","affiliation":[{"name":"Ocean University of China , , Sansha Road 1299 , ,","place":["Qingdao, PR China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,22]]},"reference":[{"key":"2026080510100813800_cit1","doi-asserted-by":"publisher","first-page":"105571","DOI":"10.1016\/j.tifs.2026.105571","article-title":"A review of tree nut\nallergens and their processing for decreasing\nallergenicity","volume":"170","author":"Ye","year":"2026","journal-title":"Trends Food Sci. 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Nutr.\nFood Res."},{"issue":"1","key":"2026080510100813800_cit5","doi-asserted-by":"publisher","first-page":"100491","DOI":"10.1016\/j.waojou.2020.100491","article-title":"Recent advances\nin the management of nut allergy","volume":"14","author":"Midun","year":"2021","journal-title":"World Allergy\nOrgan. J."},{"key":"2026080510100813800_cit6","doi-asserted-by":"publisher","first-page":"58","DOI":"10.1016\/j.molimm.2018.04.005","article-title":"Peanut allergens","volume":"100","author":"Palladino","year":"2018","journal-title":"Mol. Immunol."},{"issue":"17","key":"2026080510100813800_cit7","doi-asserted-by":"publisher","first-page":"3794","DOI":"10.1080\/10408398.2016.1175415","article-title":"Processing effects\non tree nut allergens: A review","volume":"57","author":"Vanga","year":"2017","journal-title":"Crit. Rev.\nFood Sci. 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Immunol."},{"issue":"6","key":"2026080510100813800_cit37","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0021455","article-title":"Identification of\ncritical amino\nacids in the IgE epitopes of Ric c 1 and Ric c 3 and the application\nof glutamic acid as an IgE blocker","volume":"6","author":"Deus-de-Oliveira","year":"2011","journal-title":"PLoS One"},{"issue":"6","key":"2026080510100813800_cit38","doi-asserted-by":"publisher","first-page":"499","DOI":"10.1016\/j.biochi.2005.02.011","article-title":"Molecular\nmodelling of the major\npeanut allergen Ara h 1 and other homotrimeric allergens of the cupin\nsuperfamily: a structural basis for their IgE-binding cross-reactivity","volume":"87","author":"Barre","year":"2005","journal-title":"Biochimie"},{"issue":"6","key":"2026080510100813800_cit39","doi-asserted-by":"publisher","first-page":"2990","DOI":"10.1021\/acs.jafc.2c07061","article-title":"Structure and IgE Cross-Reactivity\namong Cashew, Pistachio,\nWalnut, and Peanut Vicilin-Buried Peptides","volume":"71","author":"Foo","year":"2023","journal-title":"J. Agric. Food Chem."},{"issue":"6","key":"2026080510100813800_cit40","doi-asserted-by":"publisher","first-page":"672","DOI":"10.4168\/aair.2025.17.6.672","article-title":"Update on Tree Nut and Seed Allergies:\nPrevalence, Clinical Characteristics,\nDiagnosis, and Management","volume":"17","author":"Jeong","year":"2025","journal-title":"Allergy, Asthma Immunol.\nRes."},{"key":"2026080510100813800_cit41","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1016\/j.molimm.2020.03.017","article-title":"Epitopes\nwith similar physicochemical\nproperties contribute to cross reactivity between peanut and tree\nnuts","volume":"122","author":"Nesbit","year":"2020","journal-title":"Mol. Immunol."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c06151","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/30\/23816\/66174242\/acs.jafc.6c06151.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/30\/23816\/66174242\/acs.jafc.6c06151.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T14:31:06Z","timestamp":1785940266000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/30\/23816\/5228509\/Identification-and-Key-Amino-Acid-Analysis-of"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,22]]},"references-count":41,"journal-issue":{"issue":"30","published-online":{"date-parts":[[2026,7,22]]},"published-print":{"date-parts":[[2026,8,5]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c06151","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,22]]}}}