Tohoku J. Exp. Med., 2009, 217(3)

Lectin Activity of the Coagulation Factor VIII/von Willebrand Complex

FRANCISCA SANTIZO,¹ EDGAR ZENTENO,² SOCORRO PINA-CANSECO,³ PEDRO HERNANDEZ-CRUZ,³ MARGARITO MARTÍNEZ CRUZ,¹ LAURA PEREZ-CAMPOS MAYORAL,³ EDUARDO PÉREZ-CAMPOS^1,3 and RUTH MARTÍNEZ-CRUZ³

¹Biochemistry and Immunology Unit of Technological Institute of Oaxaca, Oaxaca, Mexico
²Department of Biochemistry, School of Medicine, National Autonomous University of Mexico, Mexico D.F., Mexico
³Medical and Biological Research Center, CICIMEBIO, School of Medicine at the Benito Juarez Autonomous University of Oaxaca, UABJO, Oaxaca, Mexico

The human coagulation factor VIII (FVIII) is essential in the intrinsic pathway of blood coagulation and circulates mainly as a non-covalently bound complex with the von Willebrand factor (VWF). This complex (FVIII/VWF) protects FVIII from degradation and cellular uptake, although no biological role has been identified yet for this complex. The FVIII/VWF complex was purified from a healthy donor's plasma by affinity chromatography on a Sepharose 4B-Concanavalin A column and was used to determine its capability to interact with erythrocytes and platelets. The purified FVIII/VWF complex at 6.0 and 12 μg/ml agglutinates rabbit and bovine erythrocytes, and showed negative agglutination with erythrocytes from other species including human ABO. Treatment of erythrocytes with Clostridium perfringens sialidase or trypsin increased four-fold the activity toward rabbit erythrocytes and positive agglutination for human A and B erythrocytes, suggesting the presence of FVIII/VWF-cryptic receptors in these erythrocytes. Goat, pig, or human O erythrocytes were not agglutinated even after enzymatic treatment. Fucose or N-acetyl-glucosamine (GlcNAc), at 10 mM, inhibited agglutinating activity of the complex with rabbit, human A and B erythrocytes, whereas galactose and N-acetyl-galactosamine, even at 200 mM, showed no effect on the complex activity. The FVIII/VWF complex, at 1.5 μg/200,000 platelets, significantly decreased platelet aggregation (p < 0.001) when compared with the effect of platelet-rich plasma; this effect was inhibited with 15 mM GlcNAc or fucose. ELISA assays on FVIII/VWF coated polystyrene plates confirmed specific binding to fucose- or biotinylated GlcNAc-dextran derivatives. We therefore propose that the FVIII/VWF complex possesses lectin activity.

keywords —— FVIII/VWF complex; coagulation factor; lectin; fucose; N-acetyl-D-glucosamine.

© 2009 Tohoku University Medical Press

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Tohoku J. Exp. Med., 2009, 217, 209-215

Correspondence: R. Martínez-Cruz, CICIMEBIO, School of Medicine, Benito Juarez Autonomous University of Oaxaca, Oaxaca 68020 Mexico.

e-mail: perezcamposcicimebio@yahoo.es, laboratory@prodigy.net.mx