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Inborn genetic variants in\n                    <jats:italic>CTRC<\/jats:italic>\n                    increase CP risk by various mechanisms that cause loss of CTRC function. Here, we investigated the functional defect of variant c.719G&gt;A (p.R240Q), which was identified in a CP case from Slovakia. CTRC secretion was analyzed using transfected HEK 293T cells. Purified wild-type and p.R240Q variant CTRC were used to determine enzyme kinetic parameters on a peptide substrate and to assess degradation of bovine \u03b2-casein and human cationic trypsinogen. Autoactivation of trypsinogen in the absence and presence of CTRC was measured. Our results indicated that the p.R240Q variant neutralized a charged amino acid that contributes to the positive electrostatic surface potential around the CTRC substrate binding site. Variant p.R240Q had no effect on CTRC secretion from transfected cells. Purified wild-type and p.R240Q enzymes cleaved a peptide substrate with comparable kinetics; while variant p.R240Q digested \u03b2-casein 2.1-fold better than wild-type CTRC. Remarkably, variant p.R240Q was 4.5-fold less effective in trypsinogen degradation and suppression of trypsinogen autoactivation, relative to the wild-type CTRC enzyme. We conclude that\u00a0\n                    <jats:italic>CTRC<\/jats:italic>\n                    variant p.R240Q increases CP risk by selectively impairing trypsinogen degradation through disruption of long-range electrostatic interactions required for efficient substrate binding. This unique case reinforces the notion that functional analysis of natural\n                    <jats:italic>CTRC<\/jats:italic>\n                    variants should always include the pathologically relevant substrate, trypsinogen.\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-40633-0","type":"journal-article","created":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T17:35:08Z","timestamp":1771954508000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Pancreatitis-associated chymotrypsin C (CTRC) variant p.R240Q selectively impairs trypsinogen degradation through disruption of long-range electrostatic interactions"],"prefix":"10.1038","volume":"16","author":[{"given":"Zolt\u00e1n Attila","family":"Nagy","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00e1t\u00e9","family":"S\u00e1ndor","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eszter","family":"Hegyi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Radovan","family":"Jur\u00ed\u010dek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriela","family":"Hr\u010dkov\u00e1","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":[[2026,2,24]]},"reference":[{"key":"40633_CR1","doi-asserted-by":"publisher","first-page":"4417","DOI":"10.3748\/wjg.v30.i41.4417","volume":"30","author":"AS Gukovskaya","year":"2024","unstructured":"Gukovskaya, A. S. et al. Trypsin in pancreatitis: The culprit, a mediator, or epiphenomenon?. World J. Gastroenterol. 30, 4417\u20134438 (2024).","journal-title":"World J. Gastroenterol."},{"key":"40633_CR2","doi-asserted-by":"publisher","first-page":"G87","DOI":"10.1152\/ajpgi.00152.2025","volume":"330","author":"A Geisz-Fremy","year":"2026","unstructured":"Geisz-Fremy, A. Pathologically relevant trypsinogen activation in pancreatitis. Am. J. Physiol. Gastrointest. Liver Physiol. 330, G87\u2013G97 (2026).","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"40633_CR3","doi-asserted-by":"publisher","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. 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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":"40633_CR7","doi-asserted-by":"publisher","first-page":"4283","DOI":"10.1111\/febs.12018","volume":"279","author":"A Szab\u00f3","year":"2012","unstructured":"Szab\u00f3, A. & Sahin-T\u00f3th, M. Determinants of chymotrypsin C cleavage specificity in the calcium-binding loop of human cationic trypsinogen. FEBS J. 279, 4283\u20134292 (2012).","journal-title":"FEBS J."},{"key":"40633_CR8","doi-asserted-by":"publisher","first-page":"4753","DOI":"10.1074\/jbc.M113.538884","volume":"289","author":"A Szab\u00f3","year":"2014","unstructured":"Szab\u00f3, A., Radisky, E. S. & Sahin-T\u00f3th, M. Zymogen activation confers thermodynamic stability on a key peptide bond and protects human cationic trypsin from degradation. J. Biol. Chem. 289, 4753\u201345761 (2014).","journal-title":"J. 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