{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:12:25Z","timestamp":1790208745163,"version":"4.1.0"},"reference-count":40,"publisher":"National Academy of Sciences","issue":"28","license":[{"start":{"date-parts":[[2024,7,3]],"date-time":"2024-07-03T00:00:00Z","timestamp":1719964800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"HHS | National Institutes of Health","doi-asserted-by":"publisher","award":["R01 DK063158"],"award-info":[{"award-number":["R01 DK063158"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2024,7,9]]},"abstract":"<jats:p>\n                    Mammalian transglutaminases, a family of Ca\n                    <jats:sup>2+<\/jats:sup>\n                    -dependent proteins, are implicated in a variety of diseases. For example, celiac disease (CeD) is an autoimmune disorder whose pathogenesis requires transglutaminase 2 (TG2) to deamidate select glutamine residues in diet-derived gluten peptides. Deamidation involves the formation of transient \u03b3-glutamyl thioester intermediates. Recent studies have revealed that in addition to the deamidated gluten peptides themselves, their corresponding thioester intermediates are also pathogenically relevant. A mechanistic understanding of this relevance is hindered by the absence of any structure of Ca\n                    <jats:sup>2+<\/jats:sup>\n                    -bound TG2. We report the X-ray crystallographic structure of human TG2 bound to an inhibitory gluten peptidomimetic and two Ca\n                    <jats:sup>2+<\/jats:sup>\n                    ions in sites previously designated as S1 and S3. Together with additional structure-guided experiments, this structure provides a mechanistic explanation for how S1 regulates formation of an inhibitory disulfide bond in TG2, while also establishing that S3 is essential for \u03b3-glutamyl thioester formation. Furthermore, our crystallographic findings and associated analyses have revealed that i) two interacting residues, H305 and E363, play a critical role in resolving the thioester intermediate into an isopeptide bond (transamidation) but not in thioester hydrolysis (deamidation); and ii) residues N333 and K176 stabilize preferred TG2 substrates and inhibitors via hydrogen bonding to nonreactive backbone atoms. Overall, the intermediate-state conformer of TG2 reported here represents a superior model to previously characterized conformers for both transition states of the TG2-catalyzed reaction.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2407066121","type":"journal-article","created":{"date-parts":[[2024,7,3]],"date-time":"2024-07-03T11:42:29Z","timestamp":1720006949000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":20,"title":["Structural and mechanistic analysis of Ca\n                    <sup>2+<\/sup>\n                    -dependent regulation of transglutaminase 2 activity using a Ca\n                    <sup>2+<\/sup>\n                    -bound intermediate state"],"prefix":"10.1073","volume":"121","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3063-6072","authenticated-orcid":false,"given":"Agnele S.","family":"Sewa","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Harrison A.","family":"Besser","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Stanford University, Stanford, CA 94305"},{"name":"Stanford Medical Scientist Training Program, Stanford University School of Medicine, Stanford, CA 94305"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irimpan I.","family":"Mathews","sequence":"additional","affiliation":[{"name":"Stanford Synchrotron Radiation Lightsource, Stanford Linear Accelerator Center National Accelerator Laboratory, Menlo Park, CA 94025"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6529-495X","authenticated-orcid":false,"given":"Chaitan","family":"Khosla","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Stanford University, Stanford, CA 94305"},{"name":"Department of Chemical Engineering, Stanford University, Stanford, CA 94305"},{"name":"Sarafan Chemistry, Engineering Medicine and Human Health, Stanford University, Stanford, CA 94305"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2024,7,3]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrm1014"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00019.2013"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2004.04476.x"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tips.2023.09.006"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00281-012-0305-0"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-394305-7.00001-X"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2032441"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2003-8"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2022.12.002"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi011715x"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2009.07420.x"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00726-009-0442-5"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0050327"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.097162"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M116.767988"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA117.001382"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.287490"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.1c04616"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1074129"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1006\/abio.1999.4255"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0107005"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.042454899"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2010.04.023"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioorg.2014.06.003"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)46929-3"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201305133"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-42004-z"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-9187-7_16"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M204521200"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(88)80928-1"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.biochem.8b00204"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-1024-7"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909047337"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889807021206"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909042589"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1107\/S2059798319011471"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444904019158"},{"key":"e_1_3_4_38_2","doi-asserted-by":"crossref","unstructured":"I. I. Mathews A. Sewa C. Khosla Transglutaminase 2 - Open State RCSB PDB rcsb.org\/structure\/unreleased\/9BC2 Deposited 7 April 2024.","DOI":"10.2210\/pdb9bc2\/pdb"},{"key":"e_1_3_4_39_2","doi-asserted-by":"crossref","unstructured":"I. I. Mathews A. Sewa C. Khosla Transglutaminase 2 - Alternate state RCSB PDB rcsb.org\/structure\/unreleased\/9BC3 Deposited 7 April 2024.","DOI":"10.2210\/pdb9bc3\/pdb"},{"key":"e_1_3_4_40_2","doi-asserted-by":"crossref","unstructured":"I. I. Mathews A. Sewa C. Khosla Transglutaminase 2 - Intermediate State RCSB PDB rcsb.org\/structure\/unreleased\/9BC4 Deposited 7 April 2024.","DOI":"10.2210\/pdb9bc3\/pdb"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2407066121","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,7,3]],"date-time":"2024-07-03T11:43:32Z","timestamp":1720007012000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2407066121"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,7,3]]},"references-count":40,"journal-issue":{"issue":"28","published-print":{"date-parts":[[2024,7,9]]}},"alternative-id":["10.1073\/pnas.2407066121"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2407066121","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,7,3]]},"assertion":[{"value":"2024-04-08","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-06-07","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-07-03","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2407066121"}}