@article {1623545,
	title = {Single-molecule imaging of von Willebrand factor reveals tension-dependent self-association},
	journal = {Blood},
	volume = {138},
	number = {23},
	year = {2021},
	pages = {2425-2434},
	abstract = {Von Willebrand factor (VWF) is an ultra-long concatemeric protein important in hemostasis and thrombosis. VWF molecules can associate with other VWF molecules, but little is known about the mechanism. Hydrodynamic drag exerts tensile force on surface-tethered VWF that extends it and is maximal at the tether point and declines linearly to zero at the downstream, free end. Using single-molecule fluorescence microscopy, we directly visualize the kinetics of binding of free VWF in flow to surface-tethered single VWF molecules and show that self-association requires elongation of tethered VWF and that association increases with tension in tethered VWF, reaches half maximum at a characteristic tension of ~10 pN, and plateaus above ~ 25 pN. Association is reversible and hence noncovalent; a sharp decrease in shear flow results in rapid dissociation of bound VWF. Tethered, primary VWF molecules can recruit more than their own mass of secondary VWF from the flow stream. Kinetics show that instead of accelerating, the rate of accumulation decreases with time, revealing an inherently self-limiting self-association mechanism. We propose that this may be because multiple tether points between secondary and primary VWF result in lower tension on the secondary VWF, which shields more highly tensioned primary VWF from further association. GPIbα binding and VWF self-association occur in the same region of high tension in tethered VWF concatemers; however, the half-maximal tension required for activation of GPIbα is higher, suggesting differences in molecular mechanisms. These results have important implications for the mechanism of platelet plug formation in hemostasis and thrombosis.},
	url = {https://pubmed.ncbi.nlm.nih.gov/34882208/},
	author = {Fu, Hongxia and Jiang, Yan and Wesley Wong and Springer, Timothy A.}
}
