{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,24]],"date-time":"2026-03-24T11:26:38Z","timestamp":1774351598455,"version":"3.50.1"},"reference-count":38,"publisher":"Georg Thieme Verlag KG","issue":"05","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Thromb Haemost"],"published-print":{"date-parts":[[2024,5]]},"abstract":"<jats:p>\n            Background\u2003Protein C (PC) is a vitamin K-dependent anticoagulant serine protease zymogen which upon activation by the thrombin\u2013thrombomodulin (TM) complex downregulates the coagulation cascade by degrading cofactors Va and VIIIa by limited proteolysis. We identified a thrombosis patient who carried a heterozygous mutation c.881G\u2009&gt;\u2009A, p.Ser252Asn (S252N) in PROC. This mutation was originally described in a report of novel mutations in patients presenting with defective PC anticoagulant activity in Paris. The research identified PC-S252N (the \u201cParis\u201d mutation) in a propositus and her family members and highlighted the critical role of Ser252 in the anticoagulation process of activated PC (APC).<\/jats:p><jats:p>\n            Material and Methods\u2003We expressed the PC-S252N mutant in mammalian cells and characterized the properties in coagulation assays to decipher the molecular basis of anticoagulant defect of this mutation.<\/jats:p><jats:p>\n            Results\u2003We demonstrated that PC-S252N had a diminished ability to TM binding, which resulted in its impaired activation by the thrombin-TM complex. However, APC-S252N exhibited a slightly stronger cleavage capacity for the chromogenic substrate. Meanwhile, the catalytic activity of APC-S252N toward FVa was significantly reduced. Sequence analysis revealed that Ser252 to Asn substitution introduced a new potential N-linked glycosylation site (252NTT254) in the catalytic domain of PC, which adversely affected both the activation process of PC and anticoagulant activity of APC.<\/jats:p><jats:p>\n            Conclusion\u2003The new N-glycosylation site (252NTT254) resulting from the mutation of Ser252 to Asn252 in PROC affects the overall structure of the protease, thereby adversely affecting the anticoagulant function of protein C. This modification has a negative impact on both TM-promoted activation of protein C and APC cleavage of FVa, ultimately leading to thrombosis in the patient.<\/jats:p>","DOI":"10.1055\/s-0043-1777133","type":"journal-article","created":{"date-parts":[[2023,11,27]],"date-time":"2023-11-27T23:53:39Z","timestamp":1701129219000},"page":"459-470","source":"Crossref","is-referenced-by-count":1,"title":["Ser252Asn Mutation Introduces a New N-Linked Glycosylation Site and Causes Type IIb Protein C Deficiency"],"prefix":"10.1055","volume":"124","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-0179-4818","authenticated-orcid":false,"given":"Shijie","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xi","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ying","family":"Song","sequence":"additional","affiliation":[{"name":"Department of Clinical Hematology and osology, Shanghai Center of Clinical Laboratory, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lei","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunli","family":"Shi","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Shanghai Center of Clinical Laboratory, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhe","family":"Lai","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qiulan","family":"Ding","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wenman","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing","family":"Dai","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuefeng","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yeling","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"194","published-online":{"date-parts":[[2023,11,27]]},"reference":[{"issue":"11","key":"ref1","doi-asserted-by":"crossref","first-page":"5532","DOI":"10.1016\/S0021-9258(19)69233-1","article-title":"Functional properties of an endothelial cell cofactor for thrombin-catalyzed activation of protein C","volume":"256","author":"W G Owen","year":"1981","journal-title":"J Biol Chem"},{"issue":"24","key":"ref2","doi-asserted-by":"crossref","first-page":"6822","DOI":"10.1002\/j.1460-2075.1996.tb01073.x","article-title":"The 2.8 A crystal structure of Gla-domainless activated protein C","volume":"15","author":"T Mather","year":"1996","journal-title":"EMBO J"},{"issue":"S 01","key":"ref3","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1111\/j.1538-7836.2007.02491.x","article-title":"Activated protein C","volume":"5","author":"J H Griffin","year":"2007","journal-title":"J Thromb Haemost"},{"issue":"15","key":"ref4","doi-asserted-by":"crossref","first-page":"9778","DOI":"10.1016\/S0021-9258(18)92888-7","article-title":"Glycosylation of human protein C affects its secretion, processing, functional activities, and activation by thrombin","volume":"266","author":"B W Grinnell","year":"1991","journal-title":"J Biol Chem"},{"issue":"30","key":"ref5","doi-asserted-by":"crossref","first-page":"20139","DOI":"10.1016\/S0021-9258(18)54901-2","article-title":"Activated protein C-catalyzed inactivation of human factor VIII and factor VIIIa. Identification of cleavage sites and correlation of proteolysis with cofactor activity","volume":"266","author":"P J Fay","year":"1991","journal-title":"J Biol Chem"},{"issue":"02","key":"ref6","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1006\/bcmd.2002.0558","article-title":"Activated protein C-dependent and -independent anticoagulant activities of protein S have different structural requirements","volume":"29","author":"M J Heeb","year":"2002","journal-title":"Blood Cells Mol Dis"},{"issue":"05","key":"ref7","doi-asserted-by":"crossref","first-page":"3151","DOI":"10.1016\/S0021-9258(17)41841-2","article-title":"Mutation of Glu-80\u2013>Lys results in a protein C mutant that no longer requires Ca2+ for rapid activation by the thrombin-thrombomodulin complex","volume":"269","author":"A R Rezaie","year":"1994","journal-title":"J Biol Chem"},{"issue":"42","key":"ref8","doi-asserted-by":"crossref","first-page":"26486","DOI":"10.1016\/S0021-9258(18)47220-1","article-title":"Identification, cloning, and regulation of a novel endothelial cell protein C\/activated protein C receptor","volume":"269","author":"K Fukudome","year":"1994","journal-title":"J Biol Chem"},{"issue":"07","key":"ref9","doi-asserted-by":"crossref","first-page":"1495","DOI":"10.1046\/j.1538-7836.2003.00300.x","article-title":"Specificity of coagulation factor signaling","volume":"1","author":"W Ruf","year":"2003","journal-title":"J Thromb Haemost"},{"issue":"08","key":"ref10","doi-asserted-by":"crossref","first-page":"3161","DOI":"10.1182\/blood-2006-09-003004","article-title":"The cytoprotective protein C pathway","volume":"109","author":"L O Mosnier","year":"2007","journal-title":"Blood"},{"issue":"19","key":"ref11","doi-asserted-by":"crossref","first-page":"2898","DOI":"10.1182\/blood-2015-02-355974","article-title":"Activated protein C: biased for translation","volume":"125","author":"J H Griffin","year":"2015","journal-title":"Blood"},{"issue":"05","key":"ref12","doi-asserted-by":"crossref","first-page":"1370","DOI":"10.1172\/JCI110385","article-title":"Deficiency of protein C in congenital thrombotic disease","volume":"68","author":"J H Griffin","year":"1981","journal-title":"J Clin Invest"},{"issue":"06","key":"ref13","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1002\/ajh.21679","article-title":"Laboratory tests for protein C deficiency","volume":"85","author":"B Khor","year":"2010","journal-title":"Am J Hematol"},{"issue":"11","key":"ref14","doi-asserted-by":"crossref","first-page":"1337","DOI":"10.5858\/2002-126-1337-LIIDAO","article-title":"Laboratory issues in diagnosing abnormalities of protein C, thrombomodulin, and endothelial cell protein C receptor","volume":"126","author":"K Kottke-Marchant","year":"2002","journal-title":"Arch Pathol Lab Med"},{"issue":"03","key":"ref15","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1111\/bjh.16776","article-title":"Guidelines on the laboratory aspects of assays used in haemostasis and thrombosis","volume":"191","author":"P Baker","year":"2020","journal-title":"Br J Haematol"},{"issue":"04","key":"ref16","doi-asserted-by":"crossref","first-page":"1069","DOI":"10.1016\/j.jtha.2023.01.019","article-title":"Corrigendum to diagnosis and management of severe congenital protein C deficiency (SCPCD): Communication from the SSC of the ISTH [J Thromb Haemost. 2022 Jul;20(7):1735-1743]","volume":"21","author":"A Minford","year":"2023","journal-title":"J Thromb Haemost"},{"issue":"01","key":"ref17","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1182\/blood.V82.1.159.bloodjournal821159","article-title":"Five novel mutations located in exons III and IX of the protein C gene in patients presenting with defective protein C anticoagulant activity","volume":"82","author":"S Gandrille","year":"1993","journal-title":"Blood"},{"issue":"19","key":"ref18","doi-asserted-by":"crossref","first-page":"6149","DOI":"10.1021\/bi015899r","article-title":"Contribution of basic residues of the 70-80-loop to heparin binding and anticoagulant function of activated protein C","volume":"41","author":"L Yang","year":"2002","journal-title":"Biochemistry"},{"issue":"07","key":"ref19","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":"15","key":"ref20","doi-asserted-by":"crossref","first-page":"2428","DOI":"10.1182\/blood-2014-12-617274","article-title":"Protein C Thr315Ala variant results in gain of function but manifests as type II deficiency in diagnostic assays","volume":"125","author":"Q Ding","year":"2015","journal-title":"Blood"},{"issue":"02","key":"ref21","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1016\/S0021-9258(17)42183-1","article-title":"Phosphatidylethanolamine incorporation into vesicles selectively enhances factor Va inactivation by activated protein C","volume":"269","author":"M D Smirnov","year":"1994","journal-title":"J Biol Chem"},{"issue":"05","key":"ref22","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":"01","key":"ref23","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1159\/000071636","article-title":"Calibrated automated thrombin generation measurement in clotting plasma","volume":"33","author":"H C Hemker","year":"2003","journal-title":"Pathophysiol Haemost Thromb"},{"issue":"S 01","key":"ref24","first-page":"61","article-title":"Coagulation laboratory testing in patients treated with argatroban","volume":"25","author":"J M Walenga","year":"1999","journal-title":"Semin Thromb Hemost"},{"key":"ref25","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1182\/asheducation.V2012.1.460.3798662","article-title":"Coagulation assays and anticoagulant monitoring","volume":"2012","author":"D M Funk","year":"2012","journal-title":"Hematology (Am Soc Hematol Educ Program)"},{"issue":"04","key":"ref26","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1055\/s-0038-1653854","article-title":"Apparent heterozygous type II protein C deficiency caused by the factor V 506 Arg to Gln mutation","volume":"73","author":"H Ireland","year":"1995","journal-title":"Thromb Haemost"},{"issue":"02","key":"ref27","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1111\/j.1365-2141.2009.08022.x","article-title":"Clinical guidelines for testing for heritable thrombophilia","volume":"149","author":"T Baglin","year":"2010","journal-title":"Br J Haematol"},{"issue":"03","key":"ref28","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1182\/blood.V73.3.712.712","article-title":"The clinical spectrum of heterozygous protein C deficiency in a large New England kindred","volume":"73","author":"E G Bovill","year":"1989","journal-title":"Blood"},{"issue":"01","key":"ref29","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1160\/TH08-11-0763","article-title":"VTE recurrence in patients with inherited deficiencies of natural anticoagulants","volume":"101","author":"F Dentali","year":"2009","journal-title":"Thromb Haemost"},{"issue":"07","key":"ref30","doi-asserted-by":"crossref","first-page":"729","DOI":"10.1001\/archinte.166.7.729","article-title":"Risk of recurrent venous thromboembolism in patients with common thrombophilia: a systematic review","volume":"166","author":"W K Ho","year":"2006","journal-title":"Arch Intern Med"},{"key":"ref31","doi-asserted-by":"crossref","first-page":"1076029620912028","DOI":"10.1177\/1076029620912028","article-title":"Laboratory limitations of excluding hereditary protein C deficiency by chromogenic assay: discrepancies of phenotype and genotype","volume":"26","author":"H Seidel","year":"2020","journal-title":"Clin Appl Thromb Hemost"},{"issue":"06","key":"ref32","doi-asserted-by":"crossref","first-page":"2101","DOI":"10.1182\/blood.V88.6.2101.bloodjournal8862101","article-title":"Compound heterozygous protein C deficiency resulting in the presence of only the beta-form of protein C in plasma","volume":"88","author":"P Simioni","year":"1996","journal-title":"Blood"},{"issue":"06","key":"ref33","doi-asserted-by":"crossref","first-page":"1712","DOI":"10.3324\/haematol.2019.227033","article-title":"Ile73Asn mutation in protein C introduces a new N-linked glycosylation site on the first EGF-domain of protein C and causes thrombosis","volume":"105","author":"Y Lu","year":"2020","journal-title":"Haematologica"},{"issue":"04","key":"ref34","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1002\/pro.5560030407","article-title":"Models of the serine protease domain of the human antithrombotic plasma factor activated protein C and its zymogen","volume":"3","author":"C L Fisher","year":"1994","journal-title":"Protein Sci"},{"issue":"37","key":"ref35","doi-asserted-by":"crossref","first-page":"22285","DOI":"10.1074\/jbc.271.37.22285","article-title":"Mutation of protease domain residues Lys37-39 in human protein C inhibits activation by the thrombomodulin-thrombin complex without affecting activation by free thrombin","volume":"271","author":"B Gerlitz","year":"1996","journal-title":"J Biol Chem"},{"issue":"02","key":"ref36","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1002\/(SICI)1097-0134(19990501)35:2<218::AID-PROT8>3.0.CO;2-2","article-title":"Probing the activation of protein C by the thrombin-thrombomodulin complex using structural analysis, site-directed mutagenesis, and computer modeling","volume":"35","author":"K E Knobe","year":"1999","journal-title":"Proteins"},{"issue":"02","key":"ref37","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1182\/blood.V96.2.585.014k30_585_593","article-title":"The autolysis loop of activated protein C interacts with factor Va and differentiates between the Arg506 and Arg306 cleavage sites","volume":"96","author":"A J Gale","year":"2000","journal-title":"Blood"},{"issue":"25","key":"ref38","doi-asserted-by":"crossref","first-page":"23105","DOI":"10.1074\/jbc.M103138200","article-title":"Secondary substrate-binding exosite in the serine protease domain of activated protein C important for cleavage at Arg-506 but not at Arg-306 in factor Va","volume":"276","author":"U Friedrich","year":"2001","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\/s-0043-1777133.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,11,5]],"date-time":"2024-11-05T12:43:46Z","timestamp":1730810626000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.thieme-connect.de\/DOI\/DOI?10.1055\/s-0043-1777133"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,27]]},"references-count":38,"journal-issue":{"issue":"05","published-online":{"date-parts":[[2024,4,23]]},"published-print":{"date-parts":[[2024,5]]}},"URL":"https:\/\/doi.org\/10.1055\/s-0043-1777133","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":[[2023,11,27]]}}}