{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T07:31:47Z","timestamp":1782804707342,"version":"3.54.5"},"reference-count":32,"publisher":"Georg Thieme Verlag KG","issue":"04","funder":[{"name":"General Program of National Natural Science Foundation of China","award":["81870107"],"award-info":[{"award-number":["81870107"]}]},{"name":"General Program of National Natural Science Foundation of China","award":["81970127"],"award-info":[{"award-number":["81970127"]}]},{"name":"General Program of National Natural Science Foundation of China","award":["82070137"],"award-info":[{"award-number":["82070137"]}]},{"name":"Shanghai Natural Science Foundation Program","award":["22ZR1439500"],"award-info":[{"award-number":["22ZR1439500"]}]},{"name":"Young Talent Support Program of College of Health Science and Technology, Shanghai Jiao Tong University School of Medicine","award":["YX20230015"],"award-info":[{"award-number":["YX20230015"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Thromb Haemost"],"published-print":{"date-parts":[[2026,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Protein C (PC) is an important physiological anticoagulant factor in humans. Activated protein C (APC) is generated from the PC zymogen through proteolytic activation by thrombin. APC inhibits thrombin generation by inactivating activated factors V and VIII via limited proteolysis. In addition to its anticoagulant function, APC also exhibits potent cytoprotective and anti-inflammatory properties. We have identified a young male with multiple-site thrombosis, who carries a heterozygous mutation c.1151A\u2009&gt;\u2009G,p.Asn384Ser(N384S) in PC. Although this mutation has been previously documented, limited functional research has been conducted to elucidate its pathogenesis.<\/jats:p>\n                  <jats:p>To elucidate the functional alternations of the N384S mutant protein C and delineate the molecular mechanism underlying thrombosis in the patient carrying this mutation.<\/jats:p>\n                  <jats:p>We expressed the recombinant PC-N384S in mammalian cells and characterized its properties in established coagulation and anti-inflammatory assay systems.<\/jats:p>\n                  <jats:p>The expression level of the PC-N384S was reduced to approximately 7% of that observed for PC-WT. The activation of PC-N384S by thrombin or thrombin\u2013thrombomodulin (TM) complex was significantly impaired, although the addition of TM exhibited a slight enhancement in the activation process. In terms of cleaving a chromogenic substrate, the catalytic efficiency reduced to approximately 50% of that observed in the wild type. In addition, in comparison with APC-WT, APC-N384S demonstrated a pronounced decline in amidolytic activity following an extended incubation period at 37\u00b0C. APC-N384S exhibited slightly impaired anticoagulant activity in either FVa inhibition assay or plasma-based assay systems. Furthermore, anti-inflammatory activity of APC-N384S was dramatically impaired as determined by evaluating the barrier-protective effect.<\/jats:p>\n                  <jats:p>The Asn384Ser mutation impairs both the anticoagulant and barrier-protective activities of protein C, thereby increasing the thrombosis risk in the heterozygous young male.<\/jats:p>","DOI":"10.1055\/a-2569-6439","type":"journal-article","created":{"date-parts":[[2025,4,18]],"date-time":"2025-04-18T18:32:37Z","timestamp":1745001157000},"page":"418-428","source":"Crossref","is-referenced-by-count":2,"title":["Asn384Ser Mutation in Protein C is Associated with Multiple-Site Thrombosis in a Young Heterozygous Male"],"prefix":"10.1055","volume":"126","author":[{"given":"Junwei","family":"Yuan","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China"},{"name":"College of Health Science and Technology, Shanghai Jiao Tong University School of Medicine, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-0179-4818","authenticated-orcid":false,"given":"Shijie","family":"Zhou","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xi","family":"Wu","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fang","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Microbial Metabolism & Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhe","family":"Lai","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiulan","family":"Ding","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenman","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuefeng","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Dai","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaobo","family":"Hu","sequence":"additional","affiliation":[{"name":"Shanghai Center for Clinical Laboratory, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yeling","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"194","published-online":{"date-parts":[[2025,4,18]]},"reference":[{"issue":"09","key":"ref1","doi-asserted-by":"crossref","first-page":"2340","DOI":"10.1182\/blood.V79.9.2340.2340","article-title":"Direct detection of activated protein C in blood from human subjects","volume":"79","author":"A Gruber","year":"1992","journal-title":"Blood"},{"issue":"06","key":"ref2","doi-asserted-by":"crossref","first-page":"1685","DOI":"10.1182\/blood.V97.6.1685","article-title":"Endothelial cell protein C receptor plays an important role in protein C activation in vivo","volume":"97","author":"F B Taylor Jr","year":"2001","journal-title":"Blood"},{"issue":"29","key":"ref3","doi-asserted-by":"crossref","first-page":"18735","DOI":"10.1016\/S0021-9258(17)32228-7","article-title":"Factor V and protein S as synergistic cofactors to activated protein C in degradation of factor VIIIa","volume":"269","author":"L Shen","year":"1994","journal-title":"J Biol Chem"},{"issue":"11","key":"ref4","doi-asserted-by":"crossref","first-page":"4708","DOI":"10.1182\/blood.V87.11.4708.bloodjournal87114708","article-title":"Comparison of activated protein C\/protein S-mediated inactivation of human factor VIII and factor 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Burnier","year":"2013","journal-title":"Blood"},{"issue":"07","key":"ref8","doi-asserted-by":"crossref","first-page":"1735","DOI":"10.1111\/jth.15732","article-title":"Diagnosis and management of severe congenital protein C deficiency (SCPCD): communication from the SSC of the ISTH","volume":"20","author":"A Minford","year":"2022","journal-title":"J Thromb Haemost"},{"issue":"8360","key":"ref9","doi-asserted-by":"crossref","first-page":"1165","DOI":"10.1016\/S0140-6736(83)91216-3","article-title":"Inherited protein C deficiency and coumarin-responsive chronic relapsing purpura fulminans in a newborn infant","volume":"2","author":"H E Branson","year":"1983","journal-title":"Lancet"},{"issue":"02","key":"ref10","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/j.braindev.2015.07.004","article-title":"Fetal hydrocephalus and neonatal stroke as the first presentation of protein C deficiency","volume":"38","author":"M Ichiyama","year":"2016","journal-title":"Brain 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Ding","year":"2015","journal-title":"Blood"},{"issue":"07","key":"ref20","doi-asserted-by":"crossref","first-page":"1358","DOI":"10.1160\/TH17-01-0043","article-title":"Gly74Ser mutation in protein C causes thrombosis due to a defect in protein S-dependent anticoagulant function","volume":"117","author":"C Chen","year":"2017","journal-title":"Thromb Haemost"},{"issue":"05","key":"ref21","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1111\/jth.14777","article-title":"Gly197Arg mutation in protein C causes recurrent thrombosis in a heterozygous carrier","volume":"18","author":"Y Lu","year":"2020","journal-title":"J Thromb Haemost"},{"issue":"04","key":"ref22","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1002\/humu.20445","article-title":"Identification and computationally-based structural interpretation of naturally occurring variants of human protein C","volume":"28","author":"E Rovida","year":"2007","journal-title":"Hum Mutat"},{"issue":"04","key":"ref23","doi-asserted-by":"crossref","first-page":"1400","DOI":"10.1182\/blood-2010-05-287623","article-title":"von Willebrand factor-mediated platelet adhesion is critical for deep vein thrombosis in mouse models","volume":"117","author":"A Brill","year":"2011","journal-title":"Blood"},{"issue":"18","key":"ref24","doi-asserted-by":"crossref","first-page":"4763","DOI":"10.1073\/pnas.1620458114","article-title":"Activation of NLRP3 inflammasome complex potentiates venous thrombosis in response to hypoxia","volume":"114","author":"N Gupta","year":"2017","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"20","key":"ref25","doi-asserted-by":"crossref","first-page":"2435","DOI":"10.1182\/blood-2016-04-710632","article-title":"Disulfide HMGB1 derived from platelets coordinates venous thrombosis in mice","volume":"128","author":"K Stark","year":"2016","journal-title":"Blood"},{"issue":"05","key":"ref26","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1182\/blood.2023022522","article-title":"Hot under the clot: venous thrombogenesis is an inflammatory process","volume":"144","author":"J Rayes","year":"2024","journal-title":"Blood"},{"issue":"10","key":"ref27","doi-asserted-by":"crossref","first-page":"1941","DOI":"10.1111\/jth.14246","article-title":"Tissue factor at the crossroad of coagulation and cell signaling","volume":"16","author":"H Zelaya","year":"2018","journal-title":"J Thromb Haemost"},{"issue":"08","key":"ref28","doi-asserted-by":"crossref","first-page":"3178","DOI":"10.1182\/blood-2004-10-3985","article-title":"Endothelial barrier protection by activated protein C through PAR1-dependent sphingosine 1-phosphate receptor-1 crossactivation","volume":"105","author":"C 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N-linked glycosylation site and causes type IIb protein C deficiency","volume":"124","author":"S Zhou","year":"2024","journal-title":"Thromb Haemost"}],"container-title":["Thrombosis and Haemostasis"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/www.thieme-connect.de\/products\/ejournals\/pdf\/10.1055\/a-2569-6439.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T23:38:03Z","timestamp":1774309083000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.thieme-connect.de\/DOI\/DOI?10.1055\/a-2569-6439"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,18]]},"references-count":32,"journal-issue":{"issue":"04","published-online":{"date-parts":[[2026,3,23]]},"published-print":{"date-parts":[[2026,4]]}},"URL":"https:\/\/doi.org\/10.1055\/a-2569-6439","archive":["Portico","CLOCKSS"],"relation":{},"ISSN":["0340-6245","2567-689X"],"issn-type":[{"value":"0340-6245","type":"print"},{"value":"2567-689X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,18]]}}}