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Table I.

Microkinetics of full-length L-selectin-, LΔcyto- and L358stop-expressing 300.19 cells interacting with GlyCAM-1 100 sites/μm2 at 0.5, 1, and 1.75 dyn/cm2



Wall shear stressa

1.75 dyn/cm2
1 dyn/cm2
0.5 dyn/cmd
L-selectin variant Full-length LΔcyto Full-length LΔcyto Full-length LΔcyto L358stop5 
Rolling fraction (%) 40 16 54 37 
Tethering fraction (%) ND ND ND ND 32 26 
Mean pause duration (ms) 20 ± 1
 (n = 104) 12 ± 1
 (n = 88) 31 ± 1
 (n = 393) 20 ± 1
 (n = 286) 61 ± 6
 (n = 76) 47 ± 4
 (n = 65) 23 ± 2
 (n = 39) 
koff (s−164.4 ± 5.4 138.7 ± 4.7 34.1 ± 2.3 61.1 ± 5.4 16.2 ± 1.6 20.9 ± 1.5 44.6 ± 3.8 
Mean step distance (μm)b
 
4 ± 0.1
 
10.2 ± 0.3
 
2.5 ± 0.1
 
4.6 ± 0.1
 
NAc
 
N.A.
 
N.A.
 


Wall shear stressa

1.75 dyn/cm2
1 dyn/cm2
0.5 dyn/cmd
L-selectin variant Full-length LΔcyto Full-length LΔcyto Full-length LΔcyto L358stop5 
Rolling fraction (%) 40 16 54 37 
Tethering fraction (%) ND ND ND ND 32 26 
Mean pause duration (ms) 20 ± 1
 (n = 104) 12 ± 1
 (n = 88) 31 ± 1
 (n = 393) 20 ± 1
 (n = 286) 61 ± 6
 (n = 76) 47 ± 4
 (n = 65) 23 ± 2
 (n = 39) 
koff (s−164.4 ± 5.4 138.7 ± 4.7 34.1 ± 2.3 61.1 ± 5.4 16.2 ± 1.6 20.9 ± 1.5 44.6 ± 3.8 
Mean step distance (μm)b
 
4 ± 0.1
 
10.2 ± 0.3
 
2.5 ± 0.1
 
4.6 ± 0.1
 
NAc
 
N.A.
 
N.A.
 
a

All measurements were performed on cell motion segments of at least 20 cells recorded at 500 frames/s.

b

Step distance was defined as the distance traveled by a rolling cell between successive tethers.

c

Not applicable, as no rolling could be detected at low shear stress.

d

The 358stop mutant did not support any cell tethering to GlyCAM-1 at shear stresses >1 dyn/cm2.

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