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Force dependence of E-selectin/sialyl Lewisx single bond dissociation

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3 Author(s)
Tees, D.F.J. ; Dept. of Chem. Eng., Pennsylvania Univ., Philadelphia, PA, USA ; Waugh, R.E. ; Hammer, D.A.

When a range of force loading rates, rf, is applied to single receptor-ligand bonds, a plot of the peak in the observed force distribution vs ln rf yields the location of transition states in the bonding potential as well as estimates of the reverse reaction rate under zero load (Merkel et al., Nature, 397:50-3, 1999). This analysis was applied to the molecules that mediate rolling adhesion of leukocytes in blood vessels by adsorbing E-selectin onto 3 μm-diam. glass fibers and coupling the selectin ligand, sialyl Lewis x (sLex) to latex microspheres. After gentle apposition of sphere and fiber by micromanipulation, the microsphere was retracted using a protocol that combines hydrodynamic and Hookean forces to apply a constant rf. The [sLex] is titrated so adhesion occurs in only ~25% of tests, implying >80% single bonds (simulations show that a few multiple bonds have little effect on parameter estimation). The peak in the force distribution was determined from video microscopy for a variety of rf. The parameters agree with those predicted to produce rolling in Monte Carlo simulations of adhesion in flow

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[Engineering in Medicine and Biology, 1999. 21st Annual Conference and the 1999 Annual Fall Meetring of the Biomedical Engineering Society] BMES/EMBS Conference, 1999. Proceedings of the First Joint  (Volume:1 )

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