Characterization of the interaction between von Willebrand factor and osteoprotegerin.
OBJECTIVE
Osteoprotegerin (OPG), a member of the tumor necrosis-factor receptor superfamily, plays an important role in bone remodeling and is also involved in vascular diseases. OPG is physically associated with von Willebrand factor (VWF), a glycoprotein involved in primary hemostasis, within the Weibel-Palade bodies (WPBs) of endothelial cells and in plasma. The present study aimed to elucidate the molecular mechanisms underlying the interaction between OPG and VWF.


RESULTS
In a solid-phase binding assay, VWF was able to bind specifically to OPG in a calcium-dependent manner. This interaction displayed strong pH dependence with optimal binding occurring at pH 6.5 and was severely impaired by chloride-ion concentrations above 40 mm. Using a series of purified VWF derivatives the functional site that supports VWF interaction with OPG was localized on its Al domain. Fluorescence microscopy on human umbilical vein endothelial cells showed co-localization of VWF and OPG in WPBs. When secretion was induced, OPG remained associated with VWF in extracellular patches of release under biochemical conditions found in blood plasma.


CONCLUSIONS
Our observations demonstrate the existence of an interactive site for OPG within the VWF A1-domain. This study established that the optimal biochemical parameters allowing a complex formation between VWF and OPG are those thought to prevail in the trans-Golgi network. These conditions would allow VWF to act as a cargo targeting OPG to WPBs. Finally, blood environments appear suitable to preserve the complex, which may participate in vascular injury, arterial calcification and inflammation.
Shahbazi
S
S
INSERM U770, Le Kremlin-Bicêtre, F-94276, and Univ Paris-Sud, Le Kremlin-Bicêtre, France.
Lenting
P J
PJ
Fribourg
C
C
Terraube
V
V
Denis
C V
CV
Christophe
O D
OD
D000595:Amino Acid Sequence; D002478:Cells, Cultured; D006801:Humans; D053244:Osteoprotegerin / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D020349:Surface Plasmon Resonance; D014841:von Willebrand Factor / Q000378:metabolism*
D053244:Osteoprotegerin; D011994:Recombinant Proteins; D014841:von Willebrand Factor