Determinants of the Formation and Activity of Factor V-phospholipid Complexes II. Molecular Properties of the Complexes

Carol L Kandall; Stephen B Shohet; T. K Akinbami; Robert W Colman

doi:10.1055/s-0038-1651340

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1975; 34(01): 271-284
DOI: 10.1055/s-0038-1651340

Original Article

Schattauer GmbH

Determinants of the Formation and Activity of Factor V-phospholipid Complexes II. Molecular Properties of the Complexes

Carol L Kandall

¹ Departments of Medicine and Pediatrics, Harvard Medical School, the Hematology Unit of the Massachusetts General Hospital, the Children’s Hospital Medical Center, Boston, Massachusetts

² University of San Francisco Departments of Medicine and Clinical Pathology, Hematology Division, San Francisco, California 94122

,

Stephen B Shohet

¹ Departments of Medicine and Pediatrics, Harvard Medical School, the Hematology Unit of the Massachusetts General Hospital, the Children’s Hospital Medical Center, Boston, Massachusetts

² University of San Francisco Departments of Medicine and Clinical Pathology, Hematology Division, San Francisco, California 94122

,

T. K Akinbami

¹ Departments of Medicine and Pediatrics, Harvard Medical School, the Hematology Unit of the Massachusetts General Hospital, the Children’s Hospital Medical Center, Boston, Massachusetts

² University of San Francisco Departments of Medicine and Clinical Pathology, Hematology Division, San Francisco, California 94122

,

Robert W Colman

¹ Departments of Medicine and Pediatrics, Harvard Medical School, the Hematology Unit of the Massachusetts General Hospital, the Children’s Hospital Medical Center, Boston, Massachusetts

² University of San Francisco Departments of Medicine and Clinical Pathology, Hematology Division, San Francisco, California 94122

› Author Affiliations

Abstract

Summary

The stability, buoyancy, and intrinsic activity of factor V-phospholipid complexes were investigated.

The decay of factor V activity in the absence of phospholipid followed first order kinetics; however, in the presence of several phospholipids a biphasic decay curve was observed. The addition of phosphatidylethanolamine to factor V produced only a small loss of activity in the first 2 minutes but decreased the subsequent rate of inactivation fourfold. A PE-factor V complex with a low bouyant density was separated from uncomplexed factor V by sucrose density ultracentrifugation. The association constant for this complex was 5 × 10⁶ M^-1 with approximately 2 moles of factor V bound per mole of lipid micelle. The isolated complex was capable of increasing prothrombin conversion 10-fold without additional phospholipid. A still lighter complex increased the rate of prothrombin conversion 18-fold.

Phosphatidyl serine produced a concentration-dependent loss of up to 95% of the factor V activity in the first 2 minutes. After ultracentrifugation on a sucrose density gradient, a PS-factor V complex of increased density was detected. This complex failed to accelerate prothrombin conversion in the intrinsic two-stage assay.

Except at very high concentrations, phosphatidylcholine did not alter the kinetics of inactivation of factor V. A factor V-phosphatidylcholine complex could not be detected after ultracentrifugation.

When added to factor V, cardiolipin (200 μg/ml), produced a rapid 50% decline in activity with a subsequent three-fold increase in the rate of inactivation. No activity was recovered after ultracentrifugation of factor V in the presence of cardiolipin.

Saturated phosphatidylethanolamine produced a concentration dependent initial loss of activity, but only a minimal increase in the subsequent rate of inactivation. At 200 μg/ml almost no light complex was detected after ultracentrifugation, but at 800 μg/ml a light complex was observed. This behavior corresponds to the ability of saturated phosphatidylethanolamine to accelerate prothrombin conversion only at very high concentrations.

Thus, phospholipids combine with factor V to form complexes which differ in their ability to accelerate prothrombin conversion. The most active species are stable lipoprotein complexes of lower buoyant density than factor V.

Full Text

References

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