Coagulation factor III (tissue factor) interaction with phospholipid vesicles induced by cadmium: characterization of the reconstituted protein-membrane complex

Steven D. Carson, William H. Konigsberg

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Coagulation factor III (tissue factor) is a membrane glycoprotein which serves as a cofactor in the proteolytic activation of factor X and factor IX by factor VIIa. Mixing of human placental factor III apoprotein with vesicles of bovine brain phospholipids does not produce significant reconstitution of factor III activity, but, when the mixture of apoprotein and vesicles is made 5 mM with CdCI2, the apoprotein is incorporated into the vesicles. Ultracentrifugation on sucrose density gradients demonstrated that the active factor III-lipid complex formed by reconstitution with vesicles had a density indistinguishable from that of the complex formed by detergent dialysis. Vesicles isolated after centrifugation were shown to range in diameter from 20 nm to over 100 nm using the electron microscope. Gel filtration showed that factor-III activity was associated with all size-classes of vesicles. The presence of factor III activity in the smaltest vesicles argues for a specific cadmium-mediated reconstitution of the apoprotein with phospholipid vesicles.

Original languageEnglish (US)
Pages (from-to)197-205
Number of pages9
JournalBioscience Reports
Volume1
Issue number3
DOIs
StatePublished - Mar 1 1981

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Thromboplastin
Cadmium
Phospholipids
Membrane Proteins
Apoproteins
Factor VIIa
Factor X
Factor IX
Dialysis
Centrifugation
Ultracentrifugation
Membrane Glycoproteins
Detergents
Gel Chromatography
Sucrose
Brain
Electron microscopes
Gels
Chemical activation
Electrons

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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abstract = "Coagulation factor III (tissue factor) is a membrane glycoprotein which serves as a cofactor in the proteolytic activation of factor X and factor IX by factor VIIa. Mixing of human placental factor III apoprotein with vesicles of bovine brain phospholipids does not produce significant reconstitution of factor III activity, but, when the mixture of apoprotein and vesicles is made 5 mM with CdCI2, the apoprotein is incorporated into the vesicles. Ultracentrifugation on sucrose density gradients demonstrated that the active factor III-lipid complex formed by reconstitution with vesicles had a density indistinguishable from that of the complex formed by detergent dialysis. Vesicles isolated after centrifugation were shown to range in diameter from 20 nm to over 100 nm using the electron microscope. Gel filtration showed that factor-III activity was associated with all size-classes of vesicles. The presence of factor III activity in the smaltest vesicles argues for a specific cadmium-mediated reconstitution of the apoprotein with phospholipid vesicles.",
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N2 - Coagulation factor III (tissue factor) is a membrane glycoprotein which serves as a cofactor in the proteolytic activation of factor X and factor IX by factor VIIa. Mixing of human placental factor III apoprotein with vesicles of bovine brain phospholipids does not produce significant reconstitution of factor III activity, but, when the mixture of apoprotein and vesicles is made 5 mM with CdCI2, the apoprotein is incorporated into the vesicles. Ultracentrifugation on sucrose density gradients demonstrated that the active factor III-lipid complex formed by reconstitution with vesicles had a density indistinguishable from that of the complex formed by detergent dialysis. Vesicles isolated after centrifugation were shown to range in diameter from 20 nm to over 100 nm using the electron microscope. Gel filtration showed that factor-III activity was associated with all size-classes of vesicles. The presence of factor III activity in the smaltest vesicles argues for a specific cadmium-mediated reconstitution of the apoprotein with phospholipid vesicles.

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