Site-directed fluorescence probing to dissect the calcium-dependent association between soluble tissue factor and factor VIIa domains
2003 (English)In: Biochimica et Biophysica Acta - Proteins and Proteomics, ISSN 1570-9639, E-ISSN 1878-1454, Vol. 1648, no 1-2, 12-16 p.Article in journal (Refereed) Published
We have used the site-directed labeling approach to study the Ca 2+-dependent docking of factor VIIa (FVIIa) to soluble tissue factor (sTF). Nine Ca2+ binding sites are located in FVIIa and even though their contribution to the overall binding between TF and FVIIa has been thoroughly studied, their importance for local protein-protein interactions within the complex has not been determined. Specifically we have monitored the association of the ?-carboxyglutamic acid (Gla), the first EGF-like (EGF1), and the protease domains (PD) of FVIIa to sTF. Our results revealed that complex formation between sTF and FVIIa during Ca2+ titration is initiated upon Ca2+ binding to EGF1, the domain containing the site of highest Ca2+ affinity. Besides we showed that a Ca 2+-loaded Gla domain is required for an optimal association of all domains of FVIIa to sTF. Ca2+ binding to the PD seems to be of some importance for the docking of this domain to sTF. © 2003 Elsevier Science B.V. All rights reserved.
Place, publisher, year, edition, pages
2003. Vol. 1648, no 1-2, 12-16 p.
Ca2+ dependence, Factor VIIa, Fluorescence, Protein-protein interaction, Tissue factor, Titration
Engineering and Technology
IdentifiersURN: urn:nbn:se:liu:diva-46620DOI: 10.1016/S1570-9639(03)00025-6OAI: oai:DiVA.org:liu-46620DiVA: diva2:267516