P-selectin binds to the D'-D3 domains of von Willebrand factor in Weibel-Palade bodies

Michaux G, Pullen TJ, Haberichter SL and Cutler DF
Source: Blood
Publication Date: (2006)
Issue: 107(10): 3922-4
Research Area:
Cancer Research/Cell Biology
Cells used in publication:
Species: human
Tissue Origin: kidney
Species: human
Tissue Origin: kidney
293 [suspension]
Species: human
Tissue Origin: kidney
293-F FreeStyleâ„¢
Species: human
Tissue Origin:
Nucleofector® I/II/2b
It has recently been shown that the ultralarge platelet-recruiting VWF strings formed immediately at exocytosis from endothelial cells may be anchored to the cell surface by an interaction with the integral membrane protein P-selectin. This finding of a new binding partner for VWF immediately raises the question of which domains of VWF bind to P-selectin. We have exploited the fact that expression of VWF in HEK293 cells triggers the formation of Weibel-Palade body-like structures that can recruit P-selectin. A suitably modified version of this assay using co-expressed truncations of VWF together with P-selectin variants in HEK2193 cells allowed us to determine which domains of VWF would recruit P-selectin within a physiologically appropriate intracellular environment. Confirming the results of such a cellular assay by conventional co-immunoprecipitation, we conclude that the lumenal domain of P-selectin interacts with the D'-D3 domains of VWF.