L-selectin is a leukocyte lectin that mediates leukocyte capture and rolling in the vasculature. The cytoplasmic domain of L-selectin has been shown to regulate leukocyte rolling. In this study, the regulatory mechanisms by which this domain controls L-selectin adhesiveness were investigated. We report that an L-selectin mutant generated by truncation of the COOH-terminal 11 residues of L-selectin tail, which impairs association with the cytoskeletal protein α-actinin, could capture leukocytes to glycoprotein L-selectin ligands under physiological shear flow. However, the conversion of initial tethers into rolling was impaired by this partial tail truncation, and was completely abolished by a further four-residue truncation of the L-selectin tail. Physical anchorage of both cell-free tail-truncated mutants within a substrate fully rescued their adhesive deficiencies. Microkinetic analysis of full-length and truncated L-selectin–mediated rolling at millisecond temporal resolution suggests that the lifetime of unstressed L-selectin tethers is unaffected by cytoplasmic tail truncation. However, cytoskeletal anchorage of L-selectin stabilizes the selectin tether by reducing the sensitivity of its dissociation rate to increasing shear forces. Low force sensitivity (reactive compliance) of tether lifetime is crucial for selectins to mediate leukocyte rolling under physiological shear stresses. This is the first demonstration that reduced reactive compliance of L-selectin tethers is regulated by cytoskeletal anchorage, in addition to intrinsic mechanical properties of the selectin–carbohydrate bond.
Skip Nav Destination
Article navigation
1 October 2001
Article|
October 01 2001
Cytoplasmic anchorage of L-selectin controls leukocyte capture and rolling by increasing the mechanical stability of the selectin tether
Oren Dwir,
Oren Dwir
1Department of Immunology, Weizmann Institute of Science, Rehovot, 76100 Israel
Search for other works by this author on:
Geoffrey S. Kansas,
Geoffrey S. Kansas
2Department of Microbiology-Immunology, Northwestern Medical School, Chicago, IL 60611
Search for other works by this author on:
Ronen Alon
Ronen Alon
1Department of Immunology, Weizmann Institute of Science, Rehovot, 76100 Israel
Search for other works by this author on:
Oren Dwir
1Department of Immunology, Weizmann Institute of Science, Rehovot, 76100 Israel
Geoffrey S. Kansas
2Department of Microbiology-Immunology, Northwestern Medical School, Chicago, IL 60611
Ronen Alon
1Department of Immunology, Weizmann Institute of Science, Rehovot, 76100 Israel
Address correspondence to Dr. Ronen Alon, Dept. of Immunology, Weizmann Institute of Science, Rehovot, 76100 Israel. Tel.: 972-8-9342482. Fax: 972-8-9344141. E-mail: [email protected]
The online version of this article contains supplemental material.
*
Abbreviations used in this paper: CB, cytochalasin B; GlyCAM-1, glycoprotein cell adhesion molecule 1; HEV, high endothelial venule; PNAd, peripheral node addressin; sLex, sialyl Lewisx.
Received:
March 12 2001
Revision Received:
August 21 2001
Accepted:
August 27 2001
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2001
J Cell Biol (2001) 155 (1): 145–156.
Article history
Received:
March 12 2001
Revision Received:
August 21 2001
Accepted:
August 27 2001
Citation
Oren Dwir, Geoffrey S. Kansas, Ronen Alon; Cytoplasmic anchorage of L-selectin controls leukocyte capture and rolling by increasing the mechanical stability of the selectin tether . J Cell Biol 1 October 2001; 155 (1): 145–156. doi: https://doi.org/10.1083/jcb.200103042
Download citation file:
Sign in
Don't already have an account? Register
Client Account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.
Sign in via your Institution
Sign in via your InstitutionEmail alerts
Advertisement
Advertisement