© The Rockefeller University Press,
0021-9525/1997//1169 $5.00
The Journal of Cell Biology, Volume 138, Number 5,
, 1997 1169-1180
The Kinetics of L-selectin Tethers and the Mechanics of Selectin-mediated Rolling
Ronen Alon,
Shuqi Chen,
Kamal D. Puri,
Erik B. Finger, and
Timothy A. Springer
The Center for Blood Research, and Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115
Two mechanisms have been proposed for regulating rolling velocities on selectins. These are (a) the intrinsic kinetics of bond dissociation, and (b) the reactive compliance, i.e., the susceptibility of the bond dissociation reaction to applied force. To determine which of these mechanisms explains the 7.5–11.5-fold faster rolling of leukocytes on L-selectin than on E- and P-selectins, we have compared the three selectins by examining the dissociation of transient tethers. We find that the intrinsic kinetics for tether bond dissociation are 7–10-fold more rapid for L-selectin than for E- and P-selectins, and are proportional to the rolling velocities through these selectins. The durations of pauses during rolling correspond to the duration of transient tethers on low density substrates. Moreover, applied force increases dissociation kinetics less for L-selectin than for E- and P-selectins, demonstrating that reactive compliance is not responsible for the faster rolling through L-selectin. Further measurements provide a biochemical and biophysical framework for understanding the molecular basis of rolling. Displacements of tethered cells during flow reversal, and measurements of the distance between successive pauses during rolling provide estimates of the length of a tether and the length of the adhesive contact zone, and suggest that rolling occurs with as few as two tethers per contact zone. Tether bond lifetime is an exponential function of the force on the bond, and the upper limit for the tether bond spring constant is of the same order of magnitude as the estimated elastic spring constant of the lectin–EGF unit. Shear uniquely enhances the rate of L-selectin transient tether formation, and conversion of tethers to rolling adhesions, providing further understanding of the shear threshold requirement for rolling through L-selectin.
Abbreviations used in this paper: HSA, human serum albumin; PNAd, peripheral node addressin; PSGL-1, P-selectin glycoprotein ligand–1.
Please address all correspondence to Timothy A. Springer, The Center for Blood Research and Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Room 251, Boston, MA 02115. Tel.: (617) 278-3200. Fax: (617) 278-3232.
R. Alon's present address is Weizmann Institute of Science, Department of Immunology, 76100 Rehovot, Israel.

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