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Biophys J, December 1999, p. 3371-3383, Vol. 77, No. 6
*Department of Biomedical Engineering, University of Virginia School of Medicine and School of Engineering and Applied Science, Charlottesville, Virginia 22908; and #Protein Design Laboratories, Inc., Fremont, California 94555 USA
Leukocyte capture and rolling on the vascular endothelium
is mediated principally by the selectin family of cell adhesion receptors. In a parallel plate flow chamber, neutrophil rolling on
purified selectins or a selectin-ligand substrate was resolved by high
speed videomicroscopy as a series of ratchet-like steps with a
characteristic time constant (Kaplanski, G., C. Farnarier, O. Tissot,
A. Pierres, A.-M. Benoliel, M. C. Alessi, S. Kaplanski, and P. Bongrand. 1993. Biophys. J. 64:1922-1933; Alon, R., D. A.
Hammer, and T. A. Springer. 1995. Nature (Lond.).
374:539-542). Under shear, neutrophil arrests due to bond
formation events were as brief as 4 ms. Pause time distributions for
neutrophils tethering on P-, E-, L-selectin, or peripheral node
addressin (PNAd) were compared at estimated single bond forces ranging
from 37 to 250 pN. Distributions of selectin mediated pause times were
fit to a first order exponential, resulting in a molecular dissociation constant (koff) for the respective selectin as a
function of force. At estimated single bond forces of 125 pN and below,
all three selectin dissociation constants fit the Bell and Hookean
spring models of force-driven bond breakage equivalently. Unstressed koff values based on the Bell model were 2.4, 2.6, 2.8, 3.8 s
1 for P-selectin, E-selectin, L-selectin,
and PNAd, respectively. Bond separation distances (reactive compliance)
were 0.39, 0.18, 1.11, 0.59 Å for P-selectin, E-selectin, L-selectin,
and PNAd, respectively. Dissociation constants for L-selectin and
P-selectin at single bond forces above 125 pN were considerably lower
than either Bell or Hookean spring model predictions, suggesting the existence of two regimes of reactive compliance. Additionally, interactions between L-selectin and its leukocyte ligand(s) were more
labile in the presence of flow than the L-selectin endothelial ligand,
PNAd, suggesting that L-selectin ligands may have different molecular
and mechanical properties. Both types of L-selectin bonds had a higher
reactive compliance than P-selectin or E-selectin bonds.
Biophys J, December 1999, p. 3371-3383, Vol. 77, No. 6
© 1999 by the Biophysical Society 0006-3495/99/12/3371/13 $2.00
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K.-C. Chang, D. F. J. Tees, and D. A. Hammer The state diagram for cell adhesion under flow: Leukocyte rolling and firm adhesion PNAS, September 22, 2000; (2000) 200240897. [Abstract] [Full Text]
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C. W. Smith Introduction: functional polarity of motile neutrophils Blood, April 15, 2000; 95(8): 2459 - 2461. [Full Text] [PDF]
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E. Evans, A. Leung, D. Hammer, and S. Simon Chemically distinct transition states govern rapid dissociation of single L-selectin bonds under force PNAS, March 27, 2001; 98(7): 3784 - 3789. [Abstract] [Full Text] [PDF]
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K.-C. Chang, D. F. J. Tees, and D. A. Hammer The state diagram for cell adhesion under flow: Leukocyte rolling and firm adhesion PNAS, October 10, 2000; 97(21): 11262 - 11267. [Abstract] [Full Text] [PDF]
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