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Published online 12 August 2002. doi:10.1083/jcb.200204041
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© The Rockefeller University Press, 0021-9525/2002/8/787 $5.00
The Journal of Cell Biology, Volume 158, Number 4, August 19, 2002 787-799

Article

 Distinct molecular and cellular contributions to stabilizing selectin-mediated rolling under flow

Tadayuki Yago1, Anne Leppänen2, Haiying Qiu2, Warren D. Marcus3, Matthias U. Nollert4, Cheng Zhu3, Richard D. Cummings2 and Rodger P. McEver1,2

1 Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation
2 Department of Biochemistry and Molecular Biology, Oklahoma Center for Medical Glycobiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104
3 Woodruff School of Mechanical Engineering and Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA 30332
4 School of Chemical Engineering and Materials Science, University of Oklahoma, Norman, OK 73109

Address correspondence to Rodger P. McEver, Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, 825 N.E. 13th St., Oklahoma City, OK 73104. Tel.: (405) 271-6480. Fax: (405) 271-3137. E-mail: rodger-mcever{at}omrf.ouhsc.edu

Leukocytes roll on selectins at nearly constant velocities over a wide range of wall shear stresses. Ligand-coupled microspheres roll faster on selectins and detach quickly as wall shear stress is increased. To examine whether the superior performance of leukocytes reflects molecular features of native ligands or cellular properties that favor selectin-mediated rolling, we coupled structurally defined selectin ligands to microspheres or K562 cells and compared their rolling on P-selectin. Microspheres bearing soluble P-selectin glycoprotein ligand (sPSGL)-1 or 2-glycosulfopeptide (GSP)-6, a GSP modeled after the NH2-terminal P-selectin–binding region of PSGL-1, rolled equivalently but unstably on P-selectin. K562 cells displaying randomly coupled 2-GSP-6 also rolled unstably. In contrast, K562 cells bearing randomly coupled sPSGL-1 or 2-GSP-6 targeted to a membrane-distal region of the presumed glycocalyx rolled more like leukocytes: rolling steps were more uniform and shear resistant, and rolling velocities tended to plateau as wall shear stress was increased. K562 cells treated with paraformaldehyde or methyl-ß-cyclodextrin before ligand coupling were less deformable and rolled unstably like microspheres. Cells treated with cytochalasin D were more deformable, further resisted detachment, and rolled slowly despite increases in wall shear stress. Thus, stable, shear-resistant rolling requires cellular properties that optimize selectin–ligand interactions.

Key Words: selectin; PSGL-1; rolling; adhesion; leukocyte


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