Identification of Galectin-1 and Galectin-3 as Novel Partners for Von Willebrand Factor

• Published in: Arteriosclerosis, thrombosis, and vascular biology Vol. 32; no. 4; pp. 894 - 901
• Main Authors: Saint-Lu, Nathalie, Oortwijn, Beatrijs D, Pegon, Julie N, Odouard, Soline, Christophe, Olivier D, de Groot, Philip G, Denis, Cécile V, Lenting, Peter J
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA American Heart Association, Inc 01.04.2012; Lippincott Williams & Wilkins
• Subjects: Animals; Atherosclerosis (general aspects, experimental research); Biological and medical sciences; Blood and lymphatic vessels; Cardiology. Vascular system; Cells, Cultured; Chlorides; Coronary heart disease; Disease Models, Animal; Diseases of the peripheral vessels. Diseases of the vena cava. Miscellaneous; Ferric Compounds; Galectin 1 - blood; Galectin 1 - deficiency; Galectin 1 - genetics; Galectin 1 - metabolism; Galectin 3 - blood; Galectin 3 - deficiency; Galectin 3 - genetics; Galectin 3 - metabolism; Heart; Human Umbilical Vein Endothelial Cells - metabolism; Humans; Immunoprecipitation; Medical sciences; Mice; Mice, Inbred C57BL; Mice, Knockout; Microscopy, Fluorescence; Platelet Adhesiveness; Protein Binding; Protein Interaction Domains and Motifs; Protein Interaction Mapping; Thrombosis - blood; Thrombosis - chemically induced; Thrombosis - genetics; Time Factors; von Willebrand Factor - genetics; von Willebrand Factor - metabolism; Vascular disease; Platelet; Hemostasis; von Willebrand factor; Atherosclerosis; galectin; Cardiovascular disease; Willebrand factor; platelets; Endothelium
• ISSN: 1079-5642; 1524-4636
• DOI: 10.1161/ATVBAHA.111.240309

OBJECTIVE—Although von Willebrand factor (VWF) is a heavily glycosylated protein, its potential to associate with glycan-binding proteins is poorly investigated. Here, we explored its interaction with the glycan-binding proteins galectin-1 and galectin-3. METHODS AND RESULTS—Immunofluorescence analysis using Duolink proximity ligation assays revealed that VWF colocalizes with galectin-1 and galectin-3 in endothelial cells, both before and after stimulation of endothelial cells. Moreover, galectin-1 was found along the typical VWF bundles that are released by endothelial cells. Galectin-1 and galectin-3 could be coprecipitated with VWF from plasma in immunoprecipitation assays, whereas plasma levels of galectin-1 and galectin-3 were significantly reduced in VWF-deficient mice. Binding studies using purified proteins confirmed that VWF could directly interact with both galectins, predominantly via its N-linked glycans. In search of the physiological relevance of the VWF-galectin interaction, we found that inhibition of galectins in in vitro perfusion assays was associated with increased VWF-platelet string formation, a phenomenon that was reproduced in galectin-1/galectin-3 double-deficient mice. These mice were also characterized by a more rapid formation of initial thrombi following ferric chloride–induced injury. CONCLUSION—We have identified galectin-1 and galectin-3 as novel partners for VWF, and these proteins may modulate VWF-mediated thrombus formation.