Clinical and functional characterization of p.Lys322stop variant in the SERPINC1 gene causing severe thrombophilia
Abstract Background Identification of mutations in the SERPINC1 has illuminated the intricate pathways underlying antithrombin (AT) deficiency. Our group identified a variation in the SERPINC1 gene (c.964 A > T, p.Lys322stop) and further investigated the mechanism of this variant causing AT defic...
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| Format: | Article |
| Language: | English |
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BMC
2024-12-01
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| Series: | Orphanet Journal of Rare Diseases |
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| Online Access: | https://doi.org/10.1186/s13023-024-03498-y |
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| _version_ | 1846112171635769344 |
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| author | Haiyue Zhang Xinyang Yue Tenglong Dai Jun Wu |
| author_facet | Haiyue Zhang Xinyang Yue Tenglong Dai Jun Wu |
| author_sort | Haiyue Zhang |
| collection | DOAJ |
| description | Abstract Background Identification of mutations in the SERPINC1 has illuminated the intricate pathways underlying antithrombin (AT) deficiency. Our group identified a variation in the SERPINC1 gene (c.964 A > T, p.Lys322stop) and further investigated the mechanism of this variant causing AT deficiency. Methods Multiple in silico tools were utilized to predict the conservation of mutations and their impact on the AT structure. The coagulation state was evaluated using the thrombin generation assay. Recombinant AT was overexpressed in HEK293T cells. Intracellular kinetics and extracellular secretion of recombinant AT-K322* were scrutinized by RT-qPCR, Western blotting, ELISA, and immunocytofluorescence. Results Analysis of conservation in silico indicated 43 out of the 143 amino acids deleted byAT-K322* in AT were highly conserved across homologous species. In vitro expression experiments showed that there was no significant difference in mRNA levels between the mutant (AT-K322*) and wild-type (AT-WT) forms of the protein. The truncated AT-K322* protein was clearly detected in cell lysates, but not in the culture medium. Conclusion AT-K322* resulted in the generation of a truncated protein, which in turn affected the secretion of AT, ultimately leading to AT deficiency. |
| format | Article |
| id | doaj-art-1c2b7586e473429ab7bf3fcff3a329bc |
| institution | Kabale University |
| issn | 1750-1172 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Orphanet Journal of Rare Diseases |
| spelling | doaj-art-1c2b7586e473429ab7bf3fcff3a329bc2024-12-22T12:47:35ZengBMCOrphanet Journal of Rare Diseases1750-11722024-12-011911810.1186/s13023-024-03498-yClinical and functional characterization of p.Lys322stop variant in the SERPINC1 gene causing severe thrombophiliaHaiyue Zhang0Xinyang Yue1Tenglong Dai2Jun Wu3Thrombosis Research Center, Beijing Jishuitan Hospital, Capital Medical UniversityDepartment of Clinical Laboratory, Peking University Fourth School of Clinical MedicineDepartment of Clinical Laboratory, Peking University Fourth School of Clinical MedicineThrombosis Research Center, Beijing Jishuitan Hospital, Capital Medical UniversityAbstract Background Identification of mutations in the SERPINC1 has illuminated the intricate pathways underlying antithrombin (AT) deficiency. Our group identified a variation in the SERPINC1 gene (c.964 A > T, p.Lys322stop) and further investigated the mechanism of this variant causing AT deficiency. Methods Multiple in silico tools were utilized to predict the conservation of mutations and their impact on the AT structure. The coagulation state was evaluated using the thrombin generation assay. Recombinant AT was overexpressed in HEK293T cells. Intracellular kinetics and extracellular secretion of recombinant AT-K322* were scrutinized by RT-qPCR, Western blotting, ELISA, and immunocytofluorescence. Results Analysis of conservation in silico indicated 43 out of the 143 amino acids deleted byAT-K322* in AT were highly conserved across homologous species. In vitro expression experiments showed that there was no significant difference in mRNA levels between the mutant (AT-K322*) and wild-type (AT-WT) forms of the protein. The truncated AT-K322* protein was clearly detected in cell lysates, but not in the culture medium. Conclusion AT-K322* resulted in the generation of a truncated protein, which in turn affected the secretion of AT, ultimately leading to AT deficiency.https://doi.org/10.1186/s13023-024-03498-ySERPINC1VTEAntithrombin deficiencyRecombinant AT protein |
| spellingShingle | Haiyue Zhang Xinyang Yue Tenglong Dai Jun Wu Clinical and functional characterization of p.Lys322stop variant in the SERPINC1 gene causing severe thrombophilia Orphanet Journal of Rare Diseases SERPINC1 VTE Antithrombin deficiency Recombinant AT protein |
| title | Clinical and functional characterization of p.Lys322stop variant in the SERPINC1 gene causing severe thrombophilia |
| title_full | Clinical and functional characterization of p.Lys322stop variant in the SERPINC1 gene causing severe thrombophilia |
| title_fullStr | Clinical and functional characterization of p.Lys322stop variant in the SERPINC1 gene causing severe thrombophilia |
| title_full_unstemmed | Clinical and functional characterization of p.Lys322stop variant in the SERPINC1 gene causing severe thrombophilia |
| title_short | Clinical and functional characterization of p.Lys322stop variant in the SERPINC1 gene causing severe thrombophilia |
| title_sort | clinical and functional characterization of p lys322stop variant in the serpinc1 gene causing severe thrombophilia |
| topic | SERPINC1 VTE Antithrombin deficiency Recombinant AT protein |
| url | https://doi.org/10.1186/s13023-024-03498-y |
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