{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,29]],"date-time":"2026-10-29T18:22:01Z","timestamp":1793298121084,"version":"4.5.0"},"reference-count":26,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,3,25]],"date-time":"2025-03-25T00:00:00Z","timestamp":1742860800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,3,25]],"date-time":"2025-03-25T00:00:00Z","timestamp":1742860800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100011019","name":"Nemzeti Kutat\u00e1si Fejleszt\u00e9si \u00e9s Innov\u00e1ci\u00f3s Hivatal","doi-asserted-by":"publisher","award":["K119386"],"award-info":[{"award-number":["K119386"]}],"id":[{"id":"10.13039\/501100011019","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011019","name":"Nemzeti Kutat\u00e1si Fejleszt\u00e9si \u00e9s Innov\u00e1ci\u00f3s Hivatal","doi-asserted-by":"publisher","award":["K135289"],"award-info":[{"award-number":["K135289"]}],"id":[{"id":"10.13039\/501100011019","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["R01DK082412"],"award-info":[{"award-number":["R01DK082412"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["R01DK117809"],"award-info":[{"award-number":["R01DK117809"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:p>The digestive protease chymotrypsin (CTR) protects the pancreas against harmful trypsin activity by promoting degradation of trypsinogen. Recently, we demonstrated that Arg236 is responsible for the higher proteolytic activity and better trypsinogen degrading capability of human CTRB2 compared to CTRB1. Introduction of Arg236 into CTRB1, which normally carries Asp236, dramatically increased degradation of human anionic trypsinogen. Here, we explored whether we could improve the activity of mouse CTRB1 by changing Gly236 to Arg (G236R mutant) and\/or by widening the substrate binding pocket (A244G mutant). We found that mutant G236R cleaved mouse anionic (T8) trypsinogen at Phe150 with 32-fold improved efficiency. In contrast, mutant G236R digested mouse cationic (T7) trypsinogen and bovine beta-casein at the same rate as wild-type mouse CTRB1. Mutation A244G reduced the activity of mouse CTRB1 against the two trypsinogen isoforms and casein. Double-mutant G236R-A244G cleaved mouse anionic (T8) trypsinogen 9.8-fold better than wild-type CTRB1 but 3.3-fold slower than single mutant G236R. Mutant G236R-A244G digested mouse cationic (T7) trypsinogen at the same rate as single-mutant A244G but degraded casein 2.3-fold slower. Taken together, the observations indicate that in the context of mouse CTRB1 the Arg236 residue increases protease activity in a substrate-specific manner, while Gly244 has an overall negative impact. The results will inform the design of preclinical mouse models with higher trypsinogen degradation ability and enhanced resilience against pancreatitis.<\/jats:p>","DOI":"10.1038\/s41598-025-94299-1","type":"journal-article","created":{"date-parts":[[2025,3,25]],"date-time":"2025-03-25T06:12:54Z","timestamp":1742883174000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Engineering mouse chymotrypsin B1 for improved trypsinogen degradation"],"prefix":"10.1038","volume":"15","author":[{"given":"Nataly C.","family":"Morales Granda","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e1s","family":"Szab\u00f3","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zsombor","family":"K\u00f6ller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00e1bor","family":"P\u00e1l","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikl\u00f3s","family":"Sahin-T\u00f3th","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,3,25]]},"reference":[{"key":"94299_CR1","doi-asserted-by":"crossref","first-page":"1692","DOI":"10.1007\/s10620-017-4601-3","volume":"62","author":"E Hegyi","year":"2017","unstructured":"Hegyi, E. & Sahin-T\u00f3th, M. Genetic risk in chronic pancreatitis: The trypsin-dependent pathway. Dig. Dis. Sci. 62, 1692\u20131701 (2017).","journal-title":"Dig. Dis. Sci."},{"key":"94299_CR2","doi-asserted-by":"crossref","first-page":"1238","DOI":"10.1111\/j.1440-1746.2011.06791.x","volume":"26","author":"J Zhou","year":"2011","unstructured":"Zhou, J. & Sahin-T\u00f3th, M. Chymotrypsin C mutations in chronic pancreatitis. J. Gastroenterol. Hepatol. 26, 1238\u20131246 (2011).","journal-title":"J. Gastroenterol. Hepatol."},{"key":"94299_CR3","doi-asserted-by":"crossref","first-page":"20701","DOI":"10.1074\/jbc.M112.360065","volume":"287","author":"A Szab\u00f3","year":"2012","unstructured":"Szab\u00f3, A. & Sahin-T\u00f3th, M. Increased activation of hereditary pancreatitis-associated human cationic trypsinogen mutants in presence of chymotrypsin C. J. Biol. Chem. 287, 20701\u201320710 (2012).","journal-title":"J. Biol. Chem."},{"key":"94299_CR4","doi-asserted-by":"crossref","DOI":"10.1172\/jci.insight.129717","volume":"4","author":"A Geisz","year":"2019","unstructured":"Geisz, A., Jancs\u00f3, Z., N\u00e9meth, B. C., Hegyi, E. & Sahin-T\u00f3th, M. Natural single-nucleotide deletion in chymotrypsinogen C gene increases severity of secretagogue-induced pancreatitis in C57BL\/6 mice. JCI Insight 4, e129717 (2019).","journal-title":"JCI Insight"},{"key":"94299_CR5","doi-asserted-by":"crossref","first-page":"1017","DOI":"10.1053\/j.gastro.2018.06.041","volume":"155","author":"Z Jancs\u00f3","year":"2018","unstructured":"Jancs\u00f3, Z., Hegyi, E. & Sahin-T\u00f3th, M. Chymotrypsin reduces the severity of secretagogue-induced pancreatitis in mice. Gastroenterology 155, 1017\u20131021 (2018).","journal-title":"Gastroenterology"},{"key":"94299_CR6","doi-asserted-by":"crossref","first-page":"11731","DOI":"10.1038\/s41598-020-68616-9","volume":"10","author":"D Mosztbacher","year":"2020","unstructured":"Mosztbacher, D., Jancs\u00f3, Z. & Sahin-T\u00f3th, M. Loss of chymotrypsin-like protease (CTRL) alters intrapancreatic protease activation but not pancreatitis severity in mice. Sci. Rep. 10, 11731 (2020).","journal-title":"Sci. Rep."},{"key":"94299_CR7","doi-asserted-by":"crossref","first-page":"G333","DOI":"10.1152\/ajpgi.00310.2023","volume":"327","author":"A Demcs\u00e1k","year":"2024","unstructured":"Demcs\u00e1k, A., Shariatzadeh, S. & Sahin-T\u00f3th, M. Secretagogue-induced pancreatitis in mice devoid of chymotrypsin. Am. J. Physiol. Gastrointest. Liver Physiol. 327, G333\u2013G344 (2024).","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"94299_CR8","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1038\/ng.2007.44","volume":"40","author":"J Rosendahl","year":"2008","unstructured":"Rosendahl, J. et al. Chymotrypsin C (CTRC) variants that diminish activity or secretion are associated with chronic pancreatitis. Nat. Genet. 40, 78\u201382 (2008).","journal-title":"Nat. Genet."},{"key":"94299_CR9","doi-asserted-by":"crossref","first-page":"1616","DOI":"10.1136\/gutjnl-2012-303090","volume":"62","author":"S Beer","year":"2013","unstructured":"Beer, S. et al. Comprehensive functional analysis of chymotrypsin C (CTRC) variants reveals distinct loss-of-function mechanisms associated with pancreatitis risk. Gut 62, 1616\u20131624 (2013).","journal-title":"Gut"},{"key":"94299_CR10","doi-asserted-by":"crossref","first-page":"1855","DOI":"10.1136\/gutjnl-2017-314454","volume":"67","author":"J Rosendahl","year":"2018","unstructured":"Rosendahl, J. et al. Genome-wide association study identifies inversion in the CTRB1-CTRB2 locus to modify risk for alcoholic and non-alcoholic chronic pancreatitis. Gut 67, 1855\u20131863 (2018).","journal-title":"Gut"},{"key":"94299_CR11","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0268859","volume":"17","author":"A Tak\u00e1ts","year":"2022","unstructured":"Tak\u00e1ts, A. et al. Risk of chronic pancreatitis in carriers of loss-of-function CTRC variants: A meta-analysis. PLoS ONE 17, e0268859 (2022).","journal-title":"PLoS ONE"},{"key":"94299_CR12","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/j.pan.2023.05.013","volume":"23","author":"G Berke","year":"2023","unstructured":"Berke, G. et al. Risk of chronic pancreatitis in carriers of the c.180C>T (p.Gly60=) CTRC variant: Case-control studies and meta-analysis. 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Proteins 93, 543\u2013554 (2025).","journal-title":"Proteins"},{"key":"94299_CR24","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","volume":"630","author":"J Abramson","year":"2024","unstructured":"Abramson, J. et al. Accurate structure prediction of biomolecular interactions with AlphaFold 3. Nature 630, 493\u2013500 (2024).","journal-title":"Nature"},{"key":"94299_CR25","doi-asserted-by":"crossref","first-page":"1249","DOI":"10.1126\/science.1546324","volume":"255","author":"L Hedstrom","year":"1992","unstructured":"Hedstrom, L., Szilagyi, L. & Rutter, W. J. Converting trypsin to chymotrypsin: the role of surface loops. Science 255, 1249\u20131253 (1992).","journal-title":"Science"},{"key":"94299_CR26","doi-asserted-by":"crossref","first-page":"1142","DOI":"10.1136\/gutjnl-2023-330788","volume":"73","author":"YC Wang","year":"2024","unstructured":"Wang, Y. C. et al. 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Gut 73, 1142\u20131155 (2024).","journal-title":"Gut"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-94299-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-94299-1","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-94299-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,25]],"date-time":"2025-03-25T06:14:30Z","timestamp":1742883270000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-94299-1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,25]]},"references-count":26,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["94299"],"URL":"https:\/\/doi.org\/10.1038\/s41598-025-94299-1","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,25]]},"assertion":[{"value":"25 January 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"12 March 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"25 March 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"10201"}}