PMID: 3768326Aug 26, 1986Paper

A domain of membrane-bound blood coagulation factor Va is located far from the phospholipid surface. A fluorescence energy transfer measurement

Biochemistry
B S IsaacsA E Johnson

Abstract

The larger subunit of blood coagulation factor Va was covalently labeled with iodoacetamido derivatives of fluorescein and rhodamine without loss of functional activity, as measured by either the one-stage clotting assay or the ability to accelerate prothrombin activation in a purified system. The spectral properties of the dyes were not altered by the presence or absence of the smaller subunit of factor Va, Ca2+, prothrombin, factor Xa, or phosphatidylcholine/phosphatidylserine (PC/PS, 4:1) vesicles. When fluorescein-labeled protein (factor VaF) was titrated with PC/PS vesicles containing either octadecylrhodamine or 5-(N-hexadecanoylamino)eosin, fluorescence energy transfer was observed between the protein-bound donor dyes and the acceptor dyes at the outer surface of the phospholipid bilayer. The extent of energy transfer correlated directly with the extent of protein binding to the vesicles monitored by light scattering. The distance of closest approach between the fluorescein on factor Va and the bilayer surface averaged 90 A for the two different acceptors. Association of factor VaF with factor Xa on the phospholipid surface reduced this separation by 7 A, but association with prothrombin did not alter the distance betwee...Continue Reading

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Citations

Nov 7, 1998·Biochimica Et Biophysica Acta·R F ZwaalE M Bevers
Feb 22, 2002·European Journal of Biochemistry·Kenji OshimaTsukasa Matsuda
Oct 1, 1989·Baillière's Clinical Haematology·R J Jenny, K G Mann
May 26, 2005·Journal of Molecular Biology·Amy E SchmidtS Paul Bajaj
Aug 28, 2004·The Journal of Biological Chemistry·Subramanian YegneswaranJohn H Griffin
Jul 23, 2013·Blood·Bernhard C LechtenbergJames A Huntington
Feb 1, 1992·Arteriosclerosis and Thrombosis : a Journal of Vascular Biology·C T Esmon
Aug 26, 2014·Biological Chemistry·Anja PomowskiJames A Huntington
Dec 1, 1991·Blood Reviews·A Gray, J Morgan

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