Fluid shear induces conformation change in human blood protein von Willebrand factor in solution.
Many of the physiological functions of von Willebrand Factor (VWF), including its binding interaction with blood platelets, are regulated by the magnitude of applied fluid/hydrodynamic stress. We applied two complementary strategies to study the effect of fluid forces on the solution structure of VWF. First, small-angle neutron scattering was used to measure protein conformation changes in response to laminar shear rates (G) up to 3000/s. Here, purified VWF was sheared in a quartz Couette cell and protein conformation was measured in real time over length scales from 2-140 nm. Second, changes in VWF structure up to 9600/s were quantified by measuring the binding of a fluorescent probe 1,1'-bis(anilino)-4-,4'-bis(naphthalene)-8,8'-disulfonate (bis-ANS) to hydrophobic pockets exposed in the sheared protein. Small angle neutron scattering studies, coupled with quantitative modeling, showed that VWF undergoes structural changes at G < 3000/s. These changes were most prominent at length scales <10 nm (scattering vector (q) range >0.6/nm). A mathematical model attributes these changes to the rearrangement of domain level features within the globular section of the protein. Studies with bis-ANS demonstrated marked increase in bis-ANS binding at G > 2300/s. Together, the data suggest that local rearrangements at the domain level may precede changes at larger-length scales that accompany exposure of protein hydrophobic pockets. Changes in VWF conformation reported here likely regulate protein function in response to fluid shear.
Singh
Indrajeet
I
Chemical and Biological Engineering, State University of New York, Buffalo, New York, USA.
Themistou
Efrosyni
E
Porcar
Lionel
L
Neelamegham
Sriram
S
D000817:Anilino Naphthalenesulfonates; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D009502:Neutrons; D011487:Protein Conformation; D012542:Scattering, Radiation; D013314:Stress, Mechanical; D014841:von Willebrand Factor / Q000737:chemistry*
D000817:Anilino Naphthalenesulfonates; D014841:von Willebrand Factor; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate)