A chimeric protein C containing the prothrombin Gla domain exhibits increased anticoagulant activity and altered phospholipid specificity.

The Journal of Biological Chemistry
M D SmirnovC T Esmon

Abstract

To determine the structural basis of phosphatidylethanolamine (PE)-dependent activated protein C (APC) activity, we prepared a chimeric molecule in which the Gla domain and hydrophobic stack of protein C were replaced with the corresponding region of prothrombin. APC inactivation of factor Va was enhanced 10-20-fold by PE. Protein S enhanced inactivation 2-fold and independently of PE. PE and protein S had little effect on the activity of the chimera. Factor Va inactivation by APC was approximately 5-fold less efficient than with the chimera on vesicles lacking PE and slightly more efficient on vesicles containing PE. The cleavage patterns of factor Va by APC and the chimera were similar, and PE enhanced the rate of Arg506 and Arg306 cleavage by APC but not the chimera. APC and the chimera bound to phosphatidylserine:phosphatidylcholine vesicles with similar affinity (Kd approximately 500 nM), and PE increased affinity 2-3-fold. Factor Va and protein S synergistically increased the affinity of APC on vesicles without PE to 140 nM and with PE to 14 nM, but they were less effective in enhancing chimera binding to either vesicle. In a factor Xa one-stage plasma clotting assay, the chimera had approximately 5 times more anticoagula...Continue Reading

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Citations

Sep 10, 2008·The Journal of Biological Chemistry·Shona HarmonJames S O'Donnell
Nov 12, 2009·Clinical and Applied Thrombosis/hemostasis : Official Journal of the International Academy of Clinical and Applied Thrombosis/Hemostasis· Zhixin Li Ming Zhao
May 23, 2012·BMC Systems Biology·Maria Cristina BravoStephen J Everse
Aug 4, 2012·Nuclear Medicine and Biology·Ming Zhao, Zhixin Li
Nov 26, 2011·Thrombosis and Haemostasis·Thomas J Cramer, Andrew J Gale
Apr 5, 2011·British Journal of Haematology·Thomas J Cramer, Andrew J Gale
Dec 10, 2009·British Journal of Haematology·Pranita P SarangiMinsoo Kim
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Jul 27, 2006·The Journal of Biological Chemistry·Roger J S PrestonDavid A Lane
Jan 17, 2008·The Journal of Biological Chemistry·Sinh TranBjörn Dahlbäck
Jan 9, 2009·The Journal of Biological Chemistry·Roger J S PrestonJames S O'Donnell
Aug 1, 2019·Journal of Thrombosis and Haemostasis : JTH·Magdalena GierulaJosefin Ahnström
Oct 31, 2000·The Journal of Biological Chemistry·O SafaN L Esmon
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Jul 24, 2020·Journal of Thrombosis and Haemostasis : JTH·Magdalena Gierula, Josefin Ahnström
Mar 10, 2000·Biochimica Et Biophysica Acta·C T Esmon
Mar 26, 1999·Biochemistry·M D SmirnovN L Esmon

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