{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T17:44:56Z","timestamp":1790703896468,"version":"4.1.0"},"reference-count":43,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,3,1]],"date-time":"2024-03-01T00:00:00Z","timestamp":1709251200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003549","name":"Hungarian Scientific Research Fund","doi-asserted-by":"publisher","award":["OTKA K-139293"],"award-info":[{"award-number":["OTKA K-139293"]}],"id":[{"id":"10.13039\/501100003549","id-type":"DOI","asserted-by":"publisher"}]},{"name":"University of Debrecen","award":["OTKA K-139293"],"award-info":[{"award-number":["OTKA K-139293"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Antithrombin (AT) is the major plasma inhibitor of thrombin (FIIa) and activated factor X (FXa), and antithrombin deficiency (ATD) is one of the most severe thrombophilic disorders. In this study, we identified nine novel AT mutations and investigated their genotype\u2013phenotype correlations. Clinical and laboratory data from patients were collected, and the nine mutant AT proteins (p.Arg14Lys, p.Cys32Tyr, p.Arg78Gly, p.Met121Arg, p.Leu245Pro, p.Leu270Argfs*14, p.Asn450Ile, p.Gly456delins_Ala_Thr and p.Pro461Thr) were expressed in HEK293 cells; then, Western blotting, N-Glycosidase F digestion, and ELISA were used to detect wild-type and mutant AT. RT-qPCR was performed to determine the expression of AT mRNA from the transfected cells. Functional studies (AT activity in the presence and in the absence of heparin and heparin-binding studies with the surface plasmon resonance method) were carried out. Mutations were also investigated by in silico methods. Type I ATD caused by altered protein synthesis (p.Cys32Tyr, p.Leu270Argfs*14, p.Asn450Ile) or secretion disorder (p.Met121Arg, p.Leu245Pro, p.Gly456delins_Ala_Thr) was proved in six mutants, while type II heparin-binding-site ATD (p.Arg78Gly) and pleiotropic-effect ATD (p.Pro461Thr) were suggested in two mutants. Finally, the pathogenic role of p.Arg14Lys was equivocal. We provided evidence to understand the pathogenic nature of novel SERPINC1 mutations through in vitro expression studies.<\/jats:p>","DOI":"10.3390\/ijms25052893","type":"journal-article","created":{"date-parts":[[2024,3,1]],"date-time":"2024-03-01T11:19:24Z","timestamp":1709291964000},"page":"2893","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Clinical and Molecular Characterization of Nine Novel Antithrombin Mutations"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7373-1459","authenticated-orcid":false,"given":"Judit","family":"K\u00e1llai","sequence":"first","affiliation":[{"name":"Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"},{"name":"HUN-REN-UD Cell Biology and Signaling Research Group, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R\u00e9ka","family":"Gindele","sequence":"additional","affiliation":[{"name":"Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krisztina","family":"P\u00e9nzes-Daku","sequence":"additional","affiliation":[{"name":"Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00e1bor","family":"Balogh","sequence":"additional","affiliation":[{"name":"Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-7628-9426","authenticated-orcid":false,"given":"R\u00e9ka","family":"Bog\u00e1ti","sequence":"additional","affiliation":[{"name":"Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B\u00e1lint","family":"B\u00e9csi","sequence":"additional","affiliation":[{"name":"Department of Medical Chemistry, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3476-794X","authenticated-orcid":false,"given":"\u00c9va","family":"Katona","sequence":"additional","affiliation":[{"name":"Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zsolt","family":"Ol\u00e1h","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology and Intensive Care, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P\u00e9ter","family":"Ilonczai","sequence":"additional","affiliation":[{"name":"J\u00f3sa Andr\u00e1s Teaching Hospital of University of Debrecen, 4400 Ny\u00edregyh\u00e1za, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zolt\u00e1n","family":"Boda","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1gnes","family":"R\u00f3na-Tas","sequence":"additional","affiliation":[{"name":"Department Medical Center of the Hungarian Defence Forces, National Hemophilia Center and Hemostasis, 1134 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L\u00e1szl\u00f3","family":"Nemes","sequence":"additional","affiliation":[{"name":"Department Medical Center of the Hungarian Defence Forces, National Hemophilia Center and Hemostasis, 1134 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Imelda","family":"Marton","sequence":"additional","affiliation":[{"name":"Department of Medicine, Albert Szent-Gy\u00f6rgyi Medical School, University of Szeged, 6720 Szeged, Hungary"},{"name":"Department of Transfusion Medicine, Albert Szent-Gy\u00f6rgyi Medical School, University of Szeged, 6720 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1483-3703","authenticated-orcid":false,"given":"Zsuzsanna","family":"Bereczky","sequence":"additional","affiliation":[{"name":"Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,3,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.thromres.2018.07.008","article-title":"The genetics of antithrombin","volume":"169","author":"Corral","year":"2018","journal-title":"Thromb. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1002\/(SICI)1098-1004(1996)7:1<7::AID-HUMU2>3.0.CO;2-B","article-title":"Molecular genetics of human antithrombin deficiency","volume":"7","author":"Perry","year":"1996","journal-title":"Hum. Mutat."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/S0268-960X(96)90034-X","article-title":"Molecular genetics of antithrombin deficiency","volume":"10","author":"Lane","year":"1996","journal-title":"Blood Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"S67","DOI":"10.1515\/CCLM.2010.368","article-title":"Antithrombin deficiency and its laboratory diagnosis","volume":"48","author":"Muszbek","year":"2010","journal-title":"Clin. Chem. Lab. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"15s","DOI":"10.1016\/0002-9343(89)80525-X","article-title":"Molecular genetics of inherited antithrombin III deficiencies","volume":"87","author":"Prochownik","year":"1989","journal-title":"Am. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1229","DOI":"10.1111\/j.1365-2516.2008.01830.x","article-title":"Inherited antithrombin deficiency: A review","volume":"14","author":"Patnaik","year":"2008","journal-title":"Haemophilia"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1097\/00001721-199206000-00012","article-title":"Antithrombin and its deficiency states","volume":"3","author":"Lane","year":"1992","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1056\/NEJM197501162920307","article-title":"Actions and interactions of antithrombin and heparin","volume":"292","author":"Rosenberg","year":"1975","journal-title":"N. Engl. J. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1326","DOI":"10.5858\/2002-126-1326-AD","article-title":"Antithrombin deficiency: Issues in laboratory diagnosis","volume":"126","author":"Duncan","year":"2002","journal-title":"Arch. Pathol. Lab. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"51058","DOI":"10.1074\/jbc.M210231200","article-title":"Intracellular accumulation of antithrombin Morioka (C95R), a novel mutation causing type I antithrombin deficiency","volume":"277","author":"Tanaka","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1016\/j.thromres.2004.08.025","article-title":"Characterization of two novel mutations of the antithrombin gene observed in Japanese thrombophilic patients","volume":"115","author":"Kurihara","year":"2005","journal-title":"Thromb. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1097\/MBC.0000000000000555","article-title":"A novel frameshift mutation leading to inherited type I antithrombin deficiency","volume":"28","author":"Mori","year":"2017","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"702","DOI":"10.1002\/ajh.20924","article-title":"Molecular basis of antithrombin deficiency in four Japanese patients with antithrombin gene abnormalities including two novel mutations","volume":"82","author":"Kyotani","year":"2007","journal-title":"Am. J. Hematol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1097\/01.mbc.0000161570.04883.25","article-title":"Characterization of molecular defect of 13387-9delG mutated antithrombin in inherited type I antithrombin deficiency","volume":"16","author":"Wang","year":"2005","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.thromres.2017.10.023","article-title":"Clinical and laboratory characteristics of antithrombin deficiencies: A large cohort study from a single diagnostic center","volume":"160","author":"Gindele","year":"2017","journal-title":"Thromb. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"404","DOI":"10.1016\/j.thromres.2014.11.022","article-title":"Prediction of the pathogenicity of antithrombin sequence variations by in silico methods","volume":"135","author":"Luxembourg","year":"2015","journal-title":"Thromb. Res."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Aguila, S., Noto, R., Luengo-Gil, G., Espin, S., Bohdan, N., de la Morena-Barrio, M.E., Penas, J., Rodenas, M.C., Vicente, V., and Corral, J. (2021). N-Glycosylation as a Tool to Study Antithrombin Secretion, Conformation, and Function. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22020516"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1182\/blood.2021014708","article-title":"Two SERPINC1 variants affecting N-glycosylation of Asn224 cause severe thrombophilia not detected by functional assays","volume":"140","author":"Suchon","year":"2022","journal-title":"Blood"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Aguila, S., Martinez-Martinez, I., Dichiara, G., Gutierrez-Gallego, R., Navarro-Fernandez, J., Vicente, V., and Corral, J. (2014). Increased N-glycosylation efficiency by generation of an aromatic sequon on N135 of antithrombin. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0114454"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4671","DOI":"10.1182\/blood.V92.12.4671","article-title":"Impaired cotranslational processing as a mechanism for type I antithrombin deficiency","volume":"92","author":"Fitches","year":"1998","journal-title":"Blood"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1055\/s-0037-1615215","article-title":"The molecular basis of antithrombin deficiency in Belgian and Dutch families","volume":"80","author":"Jochmans","year":"1998","journal-title":"Thromb. Haemost."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1002\/ajh.26413","article-title":"Molecular and clinical characterization of transient antithrombin deficiency: A new concept in congenital thrombophilia","volume":"97","author":"Minano","year":"2022","journal-title":"Am. J. Hematol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1038\/s41580-020-0250-z","article-title":"Mechanisms, regulation and functions of the unfolded protein response","volume":"21","author":"Hetz","year":"2020","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.thromres.2023.08.010","article-title":"Functional analysis of two abnormal antithrombin proteins with different intracellular kinetics","volume":"230","author":"Imai","year":"2023","journal-title":"Thromb. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1040","DOI":"10.1160\/TH16-08-0635","article-title":"Thrombotic risk according to SERPINC1 genotype in a large cohort of subjects with antithrombin inherited deficiency","volume":"117","author":"Picard","year":"2017","journal-title":"Thromb. Haemost."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1007\/BF02983246","article-title":"Five novel and four recurrent point mutations in the antithrombin gene causing venous thrombosis","volume":"78","author":"Nagaizumi","year":"2003","journal-title":"Int. J. Hematol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1358","DOI":"10.3324\/haematol.2009.015487","article-title":"Antithrombin Murcia (K241E) causing antithrombin deficiency: A possible role for altered glycosylation","volume":"95","author":"Ordonez","year":"2010","journal-title":"Haematologica"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1160\/TH10-08-0538","article-title":"Molecular basis of antithrombin deficiency","volume":"105","author":"Luxembourg","year":"2011","journal-title":"Thromb. Haemost."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2049","DOI":"10.1002\/hep.26173","article-title":"Endoplasmic reticulum polymers impair luminal protein mobility and sensitize to cellular stress in alpha1-antitrypsin deficiency","volume":"57","author":"Ordonez","year":"2013","journal-title":"Hepatology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1182\/blood.V83.1.146.146","article-title":"Antithrombin-Gly 424 Arg: A novel point mutation responsible for type 1 antithrombin deficiency and neonatal thrombosis","volume":"83","author":"Jochmans","year":"1994","journal-title":"Blood"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"931","DOI":"10.1111\/j.1538-7836.2004.00749.x","article-title":"Mutations in the shutter region of antithrombin result in formation of disulfide-linked dimers and severe venous thrombosis","volume":"2","author":"Corral","year":"2004","journal-title":"J. Thromb. Haemost."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1131","DOI":"10.1111\/jth.12606","article-title":"Role of the C-sheet in the maturation of N-glycans on antithrombin: Functional relevance of pleiotropic mutations","volume":"12","author":"Aguila","year":"2014","journal-title":"J. Thromb. Haemost."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1206","DOI":"10.1182\/blood.V79.5.1206.1206","article-title":"Antithrombin III Budapest: A single amino acid substitution (429Pro to Leu) in a region highly conserved in the serpin family","volume":"79","author":"Olds","year":"1992","journal-title":"Blood"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4216","DOI":"10.1021\/bi00067a008","article-title":"Complete nucleotide sequence of the antithrombin gene: Evidence for homologous recombination causing thrombophilia","volume":"32","author":"Olds","year":"1993","journal-title":"Biochemistry"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e161430","DOI":"10.1172\/jci.insight.161430","article-title":"Full-length antithrombin frameshift variant with aberrant C-terminus causes endoplasmic reticulum retention with a dominant-negative effect","volume":"7","author":"Toderici","year":"2022","journal-title":"JCI Insight"},{"key":"ref_36","first-page":"720","article-title":"Predicting functional effect of human missense mutations using PolyPhen-2","volume":"76","author":"Adzhubei","year":"2013","journal-title":"Curr. Protoc. Hum. Genet."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1038\/nmeth0410-248","article-title":"A method and server for predicting damaging missense mutations","volume":"7","author":"Adzhubei","year":"2010","journal-title":"Nat. Methods"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"5918","DOI":"10.1038\/s41467-020-19669-x","article-title":"Inferring the molecular and phenotypic impact of amino acid variants with MutPred2","volume":"11","author":"Pejaver","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2729","DOI":"10.1093\/bioinformatics\/btl423","article-title":"Predicting the insurgence of human genetic diseases associated to single point protein mutations with support vector machines and evolutionary information","volume":"22","author":"Capriotti","year":"2006","journal-title":"Bioinformatics"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"W452","DOI":"10.1093\/nar\/gks539","article-title":"SIFT web server: Predicting effects of amino acid substitutions on proteins","volume":"40","author":"Sim","year":"2012","journal-title":"Nucleic Acids Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1038\/nmeth0810-575","article-title":"MutationTaster evaluates disease-causing potential of sequence alterations","volume":"7","author":"Schwarz","year":"2010","journal-title":"Nat. Methods"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1038\/s41587-021-01156-3","article-title":"SignalP 6.0 predicts all five types of signal peptides using protein language models","volume":"40","author":"Teufel","year":"2022","journal-title":"Nat. Biotechnol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1797","DOI":"10.1515\/cclm-2014-0246","article-title":"Progressive chromogenic anti-factor Xa assay and its use in the classification of antithrombin deficiencies","volume":"52","author":"Kovacs","year":"2014","journal-title":"Clin. Chem. Lab. Med."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/5\/2893\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:08:01Z","timestamp":1760105281000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/5\/2893"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,1]]},"references-count":43,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2024,3]]}},"alternative-id":["ijms25052893"],"URL":"https:\/\/doi.org\/10.3390\/ijms25052893","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,3,1]]}}}