{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T01:54:46Z","timestamp":1767923686545,"version":"3.49.0"},"reference-count":51,"publisher":"Portland Press Ltd.","issue":"12","license":[{"start":{"date-parts":[[2025,6,5]],"date-time":"2025-06-05T00:00:00Z","timestamp":1749081600000},"content-version":"vor","delay-in-days":4,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000288","name":"The Royal Society","doi-asserted-by":"crossref","award":["RGS\\R1\\231147"],"award-info":[{"award-number":["RGS\\R1\\231147"]}],"id":[{"id":"10.13039\/501100000288","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000268","name":"Biotechnology and Biological Sciences Research Council","doi-asserted-by":"crossref","award":["BB\/V019635\/1"],"award-info":[{"award-number":["BB\/V019635\/1"]}],"id":[{"id":"10.13039\/501100000268","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Medical Research Council","award":["MR\/R01579\/1"],"award-info":[{"award-number":["MR\/R01579\/1"]}]}],"content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2025,6,18]]},"abstract":"<jats:p>Heme-regulated inhibitor (HRI) is one of the four mammalian kinases that phosphorylate eIF2\u03b1, facilitating a cellular response to stress through the regulation of mRNA translation. Originally identified as a heme sensor in erythroid progenitor cells, HRI has since emerged as a potential therapeutic target in both cancer and neurodegeneration. Here, we characterise two modes of HRI inhibition using structural mass spectrometry, biochemistry, and biophysics. We examined several competitive ATP-mimetic inhibitors \u2013 dabrafenib, encorafenib, and GCN2iB \u2013 and compared them with the heme-mimetic allosteric inhibitor, hemin. By combining hydrogen\u2013deuterium exchange mass spectrometry with protein models generated by AlphaFold 3, we investigated the structural basis of inhibition by dabrafenib and hemin. Our analysis revealed that hemin inhibition induces large-scale structural rearrangements in HRI, which are not observed with ATP-mimetic inhibitors. Our results suggest that HRI may be inhibited using two distinctly different modalities, which may guide future drug development.<\/jats:p>","DOI":"10.1042\/bcj20253072","type":"journal-article","created":{"date-parts":[[2025,6,5]],"date-time":"2025-06-05T15:09:36Z","timestamp":1749136176000},"page":"859-875","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Structural insights into allosteric inhibition of HRI kinase by heme binding via HDX-MS"],"prefix":"10.1042","volume":"482","author":[{"given":"Shivani","family":"Kanta","sequence":"first","affiliation":[{"name":"1Division of Cancer Research, School of Medicine, University of Dundee, Dundee DD1 9SY, U.K"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9226-1731","authenticated-orcid":false,"given":"Vanesa","family":"Vinciauskaite","sequence":"additional","affiliation":[{"name":"1Division of Cancer Research, School of Medicine, University of Dundee, Dundee DD1 9SY, U.K"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-0516-8741","authenticated-orcid":false,"given":"Graham","family":"Neill","sequence":"additional","affiliation":[{"name":"1Division of Cancer Research, School of Medicine, University of Dundee, Dundee DD1 9SY, U.K"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9733-2404","authenticated-orcid":false,"given":"Miratul M.K.","family":"Muqit","sequence":"additional","affiliation":[{"name":"2MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee DD1 5EH, U.K"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1386-4719","authenticated-orcid":false,"given":"Glenn R.","family":"Masson","sequence":"additional","affiliation":[{"name":"1Division of Cancer Research, School of Medicine, University of Dundee, Dundee DD1 9SY, U.K"}]}],"member":"288","published-online":{"date-parts":[[2025,6,17]]},"reference":[{"key":"2025061811014340600_BCJ-2025-3072C1","doi-asserted-by":"crossref","first-page":"2693","DOI":"10.1182\/blood-2006-08-041830","article-title":"Regulation of protein synthesis by the heme-regulated eIF2alpha kinase: relevance to anemias","volume":"109","author":"Chen","year":"2007","journal-title":"Blood"},{"key":"2025061811014340600_BCJ-2025-3072C2","doi-asserted-by":"crossref","DOI":"10.1182\/blood.V128.22.1037.1037","article-title":"Heme-regulated eIF2\u03b1 kinase coordinates translational repression of eIF2\u03b1P and mTORC1 signaling during Iron deficiency to mitigate ineffective erythropoiesis","volume":"128","author":"Zhang","year":"2016","journal-title":"Blood"},{"key":"2025061811014340600_BCJ-2025-3072C3","doi-asserted-by":"crossref","DOI":"10.1016\/j.jbc.2022.102451","article-title":"The heme-regulated inhibitor kinase requires dimerization for heme-sensing activity","volume":"298","author":"Ricketts","year":"2022","journal-title":"J. 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