{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T14:22:52Z","timestamp":1792592572251,"version":"4.5.0"},"reference-count":37,"publisher":"Georg Thieme Verlag KG","issue":"06","license":[{"start":{"date-parts":[[2024,9,3]],"date-time":"2024-09-03T00:00:00Z","timestamp":1725321600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"A collaborative research project from the Faculty of Science and the Faculty of Medicine Ramathibodi Hospital, Mahidol University."}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Thromb Haemost"],"published-print":{"date-parts":[[2025,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Protein C (PC) deficiency is a well-established risk factor for thromboembolism (TE), commonly manifesting in pediatric patients. This study aimed to elucidate the pathogenic mechanisms of two novel PC mutations, C238G and R189W, identified in Thai children with both venous and arterial TE.<\/jats:p><jats:p>The effects of wild-type (WT), C238G, and R189W PC variants were investigated through transient transfection of HEK293T cells. PC secretion levels were measured, and immunofluorescence analysis was performed to assess intracellular localization. ER stress-related gene expression and UPR activation were evaluated. Structural analysis was conducted to explore the significance of the C238 and R189W residue in PC functionality.<\/jats:p><jats:p>The C238G mutation led to a severe 95% reduction in PC secretion, while R189W showed a 30% decrease compared with WT. Immunofluorescence revealed that C238G-PC was predominantly retained in the ER, indicating protein misfolding. C238G-expressing cells exhibited significant upregulation of ER stress-related genes and UPR activation. In contrast, R189W resulted in only a modest increase in UPR gene expression, suggesting a less pronounced impact on protein folding and secretion. Structural analysis demonstrated the critical role of the C238 residue in maintaining PC's disulfide bond and overall conformation.<\/jats:p><jats:p>This study reveals distinct molecular mechanisms by which the C238G and R189W mutations contribute to PC deficiency and increased thrombotic risk. The findings emphasize the essential role of the C238 residue in preserving PC structure and secretion, enhancing the understanding of PC deficiency-associated TE in pediatric patients.<\/jats:p>","DOI":"10.1055\/a-2408-9529","type":"journal-article","created":{"date-parts":[[2024,9,3]],"date-time":"2024-09-03T23:40:23Z","timestamp":1725406823000},"page":"533-544","source":"Crossref","is-referenced-by-count":5,"title":["Unraveling the Molecular Pathogenesis of Protein C Deficiency\u2013Associated VTE: Insights from Protein C Mutations C238G and R189W in Thai Patients"],"prefix":"10.1055","volume":"125","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9434-2719","authenticated-orcid":false,"given":"Pansakorn","family":"Tanratana","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Faculty of Science, Mahidol University, Bangkok, Thailand"},{"name":"Ramathibodi Hemostasis and Thrombosis Center, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karnsasin","family":"Seanoon","sequence":"additional","affiliation":[{"name":"UNC Blood Research Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Panwajee","family":"Payongsri","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Praguywan","family":"Kadegasem","sequence":"additional","affiliation":[{"name":"Ramathibodi Hemostasis and Thrombosis Center, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand"},{"name":"Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ampaiwan","family":"Chuansumrit","sequence":"additional","affiliation":[{"name":"Ramathibodi Hemostasis and Thrombosis Center, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand"},{"name":"Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8039-5476","authenticated-orcid":false,"given":"Nongnuch","family":"Sirachainan","sequence":"additional","affiliation":[{"name":"Ramathibodi Hemostasis and Thrombosis Center, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand"},{"name":"Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, 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Haemost"},{"issue":"06","key":"ref4","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1016\/j.siny.2011.07.009","article-title":"Diagnosis and management of neonatal purpura fulminans","volume":"16","author":"V E Price","year":"2011","journal-title":"Semin Fetal Neonatal Med"},{"issue":"06","key":"ref5","doi-asserted-by":"crossref","first-page":"637","DOI":"10.4103\/0019-5154.169163","article-title":"Purpura fulminans due to acquired protein C deficiency","volume":"60","author":"D Mukherjee","year":"2015","journal-title":"Indian J Dermatol"},{"key":"ref6","doi-asserted-by":"crossref","first-page":"S11","DOI":"10.1186\/1471-2105-8-S1-S11","article-title":"ProCMD: a database and 3D web resource for protein C mutants","volume":"8","author":"P D'Ursi","year":"2007","journal-title":"BMC Bioinformatics"},{"issue":"05","key":"ref7","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1007\/BF02983995","article-title":"Risk factors of venous thromboembolism in thai patients","volume":"86","author":"P Angchaisuksiri","year":"2007","journal-title":"Int J Hematol"},{"issue":"02","key":"ref8","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.thromres.2009.10.016","article-title":"Protein C deficiency in Thai children with thromboembolism: a report of clinical presentations and mutation analysis","volume":"125","author":"N Sirachainan","year":"2010","journal-title":"Thromb Res"},{"issue":"02","key":"ref9","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1177\/1076029617709085","article-title":"R147W in PROC gene is a risk factor of thromboembolism in Thai children","volume":"24","author":"N Sirachainan","year":"2018","journal-title":"Clin Appl Thromb Hemost"},{"issue":"01","key":"ref10","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1002\/ajh.20043","article-title":"R147W mutation of PROC gene is common in venous thrombotic patients in Taiwanese Chinese","volume":"76","author":"W Tsay","year":"2004","journal-title":"Am J Hematol"},{"key":"ref11","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.envpol.2017.06.036","article-title":"Effects of organophosphorus flame retardant TDCPP on normal human corneal epithelial cells: implications for human health","volume":"230","author":"P Xiang","year":"2017","journal-title":"Environ Pollut"},{"issue":"05","key":"ref12","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1093\/protein\/15.5.403","article-title":"The consensus concept for thermostability engineering of proteins: further proof of concept","volume":"15","author":"M Lehmann","year":"2002","journal-title":"Protein Eng"},{"key":"ref13","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1186\/1471-2121-11-67","article-title":"Functional characterization of the protein C A267T mutation: evidence for impaired secretion due to defective intracellular transport","volume":"11","author":"L Tjeldhorn","year":"2010","journal-title":"BMC Cell 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Protein misfolding and ER stress","volume":"307","author":"J E Chambers","year":"2014","journal-title":"Am J Physiol Cell Physiol"},{"issue":"03","key":"ref20","doi-asserted-by":"crossref","first-page":"660","DOI":"10.1016\/j.bbadis.2017.12.016","article-title":"Factor VII deficiency: unveiling the cellular and molecular mechanisms underlying three model alterations of the enzyme catalytic domain","volume":"1864","author":"M E Chollet","year":"2018","journal-title":"Biochim Biophys Acta Mol Basis Dis"},{"issue":"01","key":"ref21","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1242\/dmm.013474","article-title":"Mechanisms of protein-folding diseases at a glance","volume":"7","author":"J S Valastyan","year":"2014","journal-title":"Dis Model Mech"},{"issue":"22","key":"ref22","doi-asserted-by":"crossref","first-page":"2460","DOI":"10.2174\/1568026611212220002","article-title":"Protein misfolding in disease and small molecule therapies","volume":"12","author":"C M Gomes","year":"2012","journal-title":"Curr Top Med Chem"},{"key":"ref23","doi-asserted-by":"crossref","first-page":"20","DOI":"10.3389\/fmolb.2019.00020","article-title":"Protein misfolding and ER stress in Huntington's disease","volume":"6","author":"T Shacham","year":"2019","journal-title":"Front Mol Biosci"},{"issue":"03","key":"ref24","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1038\/sj.cdd.4401840","article-title":"From acute ER stress to physiological roles of the unfolded protein response","volume":"13","author":"J Wu","year":"2006","journal-title":"Cell Death Differ"},{"issue":"01","key":"ref25","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.tcb.2003.11.001","article-title":"A trip to the ER: coping with stress","volume":"14","author":"D T Rutkowski","year":"2004","journal-title":"Trends Cell Biol"},{"issue":"01","key":"ref26","first-page":"45","article-title":"Endoplasmic reticulum stress and unfolded protein response pathways: potential for treating age-related retinal degeneration","volume":"7","author":"M Haeri","year":"2012","journal-title":"J Ophthalmic Vis Res"},{"issue":"09","key":"ref27","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1038\/nri2359","article-title":"The endoplasmic reticulum stress response in immunity and autoimmunity","volume":"8","author":"D J Todd","year":"2008","journal-title":"Nat Rev Immunol"},{"issue":"04","key":"ref28","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1038\/sj.cdd.4401373","article-title":"Roles of CHOP\/GADD153 in endoplasmic reticulum stress","volume":"11","author":"S Oyadomari","year":"2004","journal-title":"Cell Death Differ"},{"issue":"02","key":"ref29","first-page":"1000107","article-title":"Endoplasmic reticulum stress and related pathological processes","volume":"1","author":"Y Mei","year":"2013","journal-title":"J Pharmacol Biomed 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Sodium ion in the APC crystal structure is coordinated to four carbonyl groups from two separate loops","volume":"277","author":"A E Schmidt","year":"2002","journal-title":"J Biol Chem"}],"container-title":["Thrombosis and Haemostasis"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/www.thieme-connect.de\/products\/ejournals\/pdf\/10.1055\/a-2408-9529.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,5,27]],"date-time":"2025-05-27T22:30:50Z","timestamp":1748385050000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.thieme-connect.de\/DOI\/DOI?10.1055\/a-2408-9529"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,9,3]]},"references-count":37,"journal-issue":{"issue":"06","published-online":{"date-parts":[[2025,5,27]]},"published-print":{"date-parts":[[2025,6]]}},"URL":"https:\/\/doi.org\/10.1055\/a-2408-9529","archive":["Portico","CLOCKSS"],"relation":{},"ISSN":["0340-6245","2567-689X"],"issn-type":[{"value":"0340-6245","type":"print"},{"value":"2567-689X","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2024,9,3]]},"published":{"date-parts":[[2024,9,3]]}}}