{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:12:29Z","timestamp":1764288749361,"version":"3.46.0"},"reference-count":37,"publisher":"Georg Thieme Verlag KG","issue":"12","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Thromb Haemost"],"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>We previously identified a factor (F)VIII molecular defect associated with an R2159C mutation in the C1 domain (named \u201cFVIII-Ise\u201d) together with undetectable FVIII antigen (FVIII:Ag) levels measured by two-site sandwich ELISA using an anti-C2 domain alloantibody (alloAb). The patient had clinically mild hemophilia A, and his reduced FVIII:C correlated with FVIII:Ag measured by ELISA using monoclonal antibodies (mAbs) with A2 and A2\/B domain epitopes, suggesting that the R2159C mutation modified C2 domain antigenicity.<\/jats:p>\n                  <jats:p>To investigate functional and structural characteristics of the FVIII-R2159C mutant.<\/jats:p>\n                  <jats:p>ELISAs using a previous anti-C2 domain alloAb confirmed that the antigen level of recombinant FVIII-R2159C mutant prepared in BHK cells was 56% lower relative to wild-type (WT), consistent with our earlier reports. This anti-C2 domain alloAb competitively inhibited FVIII and anti-C1 domain mAb binding, indicating the involvement of specificity for C1 and C2 epitopes. The K\n                  m for FVIII-R2159C with FIXa or FX in the tenase complex was similar to that of FVIII-WT. Thrombin- and FXa-catalyzed cleavage reactions of FVIII-R2159C were similar to those of WT. The K\n                  d for FVIII-R2159C binding to phospholipids was moderately greater than for FVIII-WT, however, while there were no significant differences in von Willebrand factor binding. In silico molecular dynamic simulation analyses revealed subtle differences between FVIII-WT and FVIII-R2159C.<\/jats:p>\n                  <jats:p>The FVIII-R2159C mutation was not different from FVIII-WT in interactions with FIXa, FX, and thrombin, but reduced binding potential to phospholipids and to an anti-C1\/C2 domain alloAb was evident apparently due to subtle changes in conformational structure.<\/jats:p>","DOI":"10.1055\/a-2509-0511","type":"journal-article","created":{"date-parts":[[2025,1,21]],"date-time":"2025-01-21T19:23:48Z","timestamp":1737487428000},"page":"1216-1225","source":"Crossref","is-referenced-by-count":0,"title":["Structural Conformation and the Binding of Factor VIII R2159C (FVIII-Ise) Mutated in the C1 Domain to Phospholipid"],"prefix":"10.1055","volume":"125","author":[{"given":"Kuniyoshi","family":"Mizumachi","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9348-9815","authenticated-orcid":false,"given":"Masahiro","family":"Takeyama","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"},{"name":"Division of Hemophilia, National Hospital Organization, Osaka National Hospital, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kaoru","family":"Horiuchi","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keiji","family":"Nogami","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"194","published-online":{"date-parts":[[2025,1,21]]},"reference":[{"issue":"01","key":"ref1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1182\/blood.V76.1.1.1","article-title":"Surface-dependent reactions of the vitamin K-dependent enzyme complexes","volume":"76","author":"K G Mann","year":"1990","journal-title":"Blood"},{"issue":"5992","key":"ref2","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1038\/312337a0","article-title":"Structure of human factor VIII","volume":"312","author":"G A Vehar","year":"1984","journal-title":"Nature"},{"issue":"5992","key":"ref3","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1038\/312330a0","article-title":"Expression of active human factor VIII from recombinant DNA clones","volume":"312","author":"W I Wood","year":"1984","journal-title":"Nature"},{"issue":"21","key":"ref4","doi-asserted-by":"crossref","first-page":"10451","DOI":"10.1016\/S0021-9258(19)81537-5","article-title":"Association of the factor VIII light chain with von Willebrand factor","volume":"263","author":"P Lollar","year":"1988","journal-title":"J Biol Chem"},{"issue":"11","key":"ref5","doi-asserted-by":"crossref","first-page":"5230","DOI":"10.1016\/S0021-9258(18)60704-5","article-title":"An immunogenic region within residues Val1670-Glu1684 of the factor VIII light chain induces antibodies which inhibit binding of factor VIII to von Willebrand factor","volume":"263","author":"P A Foster","year":"1988","journal-title":"J Biol Chem"},{"issue":"15","key":"ref6","doi-asserted-by":"crossref","first-page":"11601","DOI":"10.1016\/S0021-9258(19)78167-8","article-title":"A role for the C2 domain of factor VIII in binding to von Willebrand factor","volume":"269","author":"E L Saenko","year":"1994","journal-title":"J Biol Chem"},{"issue":"03","key":"ref7","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1111\/j.1365-2141.1995.tb05374.x","article-title":"Common inhibitory effects of human anti-C2 domain inhibitor alloantibodies on factor VIII binding to von Willebrand factor","volume":"91","author":"M Shima","year":"1995","journal-title":"Br J Haematol"},{"issue":"03","key":"ref8","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1055\/s-0038-1651588","article-title":"A factor VIII neutralizing monoclonal antibody and a human inhibitor alloantibody recognizing epitopes in the C2 domain inhibit factor VIII binding to von Willebrand factor and to phosphatidylserine","volume":"69","author":"M Shima","year":"1993","journal-title":"Thromb Haemost"},{"issue":"02","key":"ref9","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1021\/bi00350a035","article-title":"Proteolytic processing of human factor VIII. 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Recommendation of the scientific subcommittee on factor VIII and factor IX of the scientific and standardization committee of the International Society on Thrombosis and Haemostasis","volume":"85","author":"G C White II","year":"2001","journal-title":"Thromb Haemost"},{"issue":"19","key":"ref14","doi-asserted-by":"crossref","first-page":"8307","DOI":"10.1073\/pnas.88.19.8307","article-title":"Molecular characterization of mild-to-moderate hemophilia A: detection of the mutation in 25 of 29 patients by denaturing gradient gel electrophoresis","volume":"88","author":"M Higuchi","year":"1991","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"05","key":"ref15","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1055\/s-0040-1721385","article-title":"Spectrum of F8 genotype and genetic impact on inhibitor development in patients with hemophilia A from multicenter cohort studies (J-HIS) in Japan","volume":"121","author":"K Shinozawa","year":"2021","journal-title":"Thromb Haemost"},{"issue":"05","key":"ref16","doi-asserted-by":"crossref","first-page":"862","DOI":"10.1055\/s-0038-1656068","article-title":"Factor VIII Ise (R2159C) in a patient with mild hemophilia A, an abnormal factor VIII with retention of function but modification of C2 epitopes","volume":"77","author":"H Suzuki","year":"1997","journal-title":"Thromb Haemost"},{"issue":"04","key":"ref17","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1021\/bi00507a028","article-title":"Phospholipid vesicle formation and transmembrane protein incorporation using octyl glucoside","volume":"20","author":"L T Mimms","year":"1981","journal-title":"Biochemistry"},{"issue":"03","key":"ref18","doi-asserted-by":"crossref","first-page":"820","DOI":"10.1021\/bi201731p","article-title":"Factor VIII light chain contains a binding site for factor X that contributes to the catalytic efficiency of factor Xase","volume":"51","author":"M 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Gla domain in the factor Xase complex","volume":"284","author":"T Soeda","year":"2009","journal-title":"J Biol Chem"},{"issue":"11","key":"ref37","doi-asserted-by":"crossref","first-page":"1954","DOI":"10.1182\/blood-2013-02-483263","article-title":"Factor VIII gene (F8) mutation and risk of inhibitor development in nonsevere hemophilia A","volume":"122","author":"C L Eckhardt","year":"2013","journal-title":"Blood"}],"container-title":["Thrombosis and Haemostasis"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/www.thieme-connect.de\/products\/ejournals\/pdf\/10.1055\/a-2509-0511.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:11:08Z","timestamp":1764288668000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.thieme-connect.de\/DOI\/DOI?10.1055\/a-2509-0511"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,21]]},"references-count":37,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2025,11,27]]},"published-print":{"date-parts":[[2025,12]]}},"URL":"https:\/\/doi.org\/10.1055\/a-2509-0511","archive":["Portico","CLOCKSS"],"relation":{},"ISSN":["0340-6245","2567-689X"],"issn-type":[{"type":"print","value":"0340-6245"},{"type":"electronic","value":"2567-689X"}],"subject":[],"published":{"date-parts":[[2025,1,21]]}}}