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SERPINC1 c.1247dupC: a novel SERPINC1 gene mutation associated with familial thrombosis results in a secretion defect
Maximilian Ruf1, Sarah Cunningham1, Alexandra Wandersee1
1Department of Transfusion Medicine and Hemostaseology, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), University Hospital Erlangen, Krankenhausstr. 12, 91054, Erlangen, Germany.
Insights
A novel SERPINC1 gene mutation, c.1247dupC, causes a frameshift leading to impaired antithrombin (AT) secretion. This molecular defect explains type I AT deficiency and associated thrombosis risk.
Area of Science:
- Genetics and Molecular Biology
- Hemostasis and Thrombosis
Background:
- Antithrombin (AT) is a key anticoagulant protein regulating hemostasis.
- Type I AT deficiency is linked to clinically significant thrombosis.
- Inherited SERPINC1 mutations are a known cause of AT deficiency.
Purpose of the Study:
- To characterize a novel SERPINC1 mutation identified in patients with type I AT deficiency.
- To investigate the cellular and molecular consequences of the identified mutation.
Main Methods:
- Genetic analysis to identify SERPINC1 mutations.
- Expression of wild-type and mutant SERPINC1 in HEK293T cells using GFP/HA tags.
- Analysis of protein localization and secretion via confocal microscopy and Western blot.
Main Results:
- A frameshift mutation (c.1247dupC) in SERPINC1 was identified, resulting in an altered amino acid sequence (p.Ser417LysfsTer48).
- The mutation affects the C-terminus of the antithrombin protein.
- Impaired secretion of the mutant AT protein was confirmed in cellular assays.
Conclusions:
- The p.Ser417LysfsTer48 mutation leads to reduced AT secretion, causing quantitative AT deficiency.
- This molecular mechanism explains the observed type I AT deficiency in the affected individuals.
- The findings highlight the importance of proper AT secretion for maintaining hemostatic balance.
Background:
Antithrombin (AT) is an important anticoagulant in hemostasis. We describe here the characterization of a novel AT mutation associated with clinically relevant thrombosis. A pair of sisters with confirmed type I AT protein deficiency was genetically analyzed on suspicion of an inherited SERPINC1 mutation. A frameshift mutation, c.1247dupC, was identified and the effect of this mutation was examined on the cellular and molecular level.
Methods:
Plasmids for the expression of wild-type (WT) and mutated SERPINC1 coding sequence (CDS) fused to green fluorescent protein (GFP) or hemagglutinin (HA) tag were transfected into HEK293T cells. Subcellular localization and secretion of the respective fusion proteins were analyzed by confocal laser scanning microscopy and Western blot.
Results:
The c.1247dupC mutation results in a frameshift in the CDS of the SERPINC1 gene and a subsequently altered amino acid sequence (p.Ser417LysfsTer48). This alteration affects the C-terminus of the AT antigen and results in impaired secretion as confirmed by GFP- and HA-tagged mutant AT analyzed in HEK293T cells.
Conclusion:
The p.Ser417LysfsTer48 mutation leads to impaired secretion, thus resulting in a quantitative AT deficiency. This is in line with the type I AT deficiency observed in the patients.
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