Coagulation factor VII Gln100 --> Arg. Amino acid substitution at the epidermal growth factor 2-protease domain interface results in severely reduced tissue factor binding and procoagulant function.

The Journal of Biological Chemistry
G Kemball-CookE G Tuddenham

Abstract

We have used recombinant mammalian expression and purification of the factor VII (FVII) variant Gln100 --> Arg (Q100RFVII) to study FVII deficiency in subjects with this mutation. Q100RFVII was secreted poorly in comparison with wild-type FVII (WTFVII) in a stable mammalian expression system, and purified variant protein was found to have undetectable clotting activity. Following activation by immobilized factor Xa, Q100RFVIIa had amidolytic activity similar to WTFVIIa in the absence of soluble tissue factor (sTF); however, unlike WTFVIIa no typical increase in activity was seen after addition of sTF. In a factor X activation assay using relipidated transmembrane truncated tissue factor (residues 1-243), Q100RFVIIa showed less than 5% of the ability of WTFVIIa to activate factor X. We performed direct binding analysis of WT and Q100RFVII/FVIIa to immobilized sTF using surface plasmon resonance, and severely reduced binding of both non-activated and activated Q100RFVII to sTF was seen, indicating a pronounced defect in tissue factor (TF) interaction with this variant. In the sTF-FVIIa crystal structure the candidate residue Gln100 is not in contact with TF but is at the epidermal growth factor 2-protease domain interface. We sug...Continue Reading

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Citations

Dec 28, 1999·Journal of Molecular Recognition : JMR·D G Myszka
Jan 4, 2001·Human Mutation·J H McVeyE G Tuddenham
Mar 27, 2002·Journal of Computational Chemistry·Lalith PereraLee G Pedersen
Aug 11, 2000·Blood Coagulation & Fibrinolysis : an International Journal in Haemostasis and Thrombosis·M SutoT Naruse
Aug 28, 2003·Journal of Thrombosis and Haemostasis : JTH·E B BachliJ H McVey
Sep 12, 2006·Journal of Thrombosis and Haemostasis : JTH·M B CallanK A High
Aug 19, 2007·European Journal of Haematology·Rajiv K PruthiElizabeth A Plumhoff
Sep 3, 2013·Haemophilia : the Official Journal of the World Federation of Hemophilia·H T T TranP A Holme
Nov 2, 1999·Journal of Structural Biology·G Kemball-CookK Harlos

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