Glycolipids support E-selectin-specific strong cell tethering under flow

Monica M. Burdick; Bruce S. Bochner; Brian E. Collins; Ronald L. Schnaar; Konstantinos Konstantopoulos

doi:10.1006/bbrc.2001.4899

Glycolipids support E-selectin-specific strong cell tethering under flow

Monica M. Burdick, Bruce S. Bochner, Brian E. Collins, Ronald L. Schnaar, Konstantinos Konstantopoulos^*

^*Corresponding author for this work

Medicine, Allergy Division

Research output: Contribution to journal › Article › peer-review

37 Scopus citations

Abstract

This study provides functional evidence that glycosphingolipids constitute ligands for E-selectin but not P-selectin. Chinese hamster ovary (CHO) cells expressing E-selectin (CHO-E) or P-selectin (CHO-P) were perfused over α2,3-sialyl Lewis X (α2,3-sLe^x) presented as the hexaosylceramide glycosphingolipid adsorbed in a monolayer containing phosphatidylcholine and cholesterol. CHO-E cells tethered extensively and formed slow, stable rolling interactions with α2,3-sLe^x glycosphingolipid but not with the comparable α2,6-sLe^x glycosphingolipid. Tethering/rolling varied with wall shear stress, selectin density, and ligand density. In contrast, α2,3-sLe^x glycosphingolipid supported only limited, fast CHO-P cell rolling. As calculated from a stochastic model of cell rolling, the step size between successive bond releases from the α2,3-sLe^x glycosphingolipid was smaller for CHO-E than CHO-P cells, whereas the opposite effect was observed for the waiting time between these events. Detachment assays revealed stronger adhesive interactions of CHO-E than CHO-P cells with α2,3-sLe^x glycosphingolipid. These findings indicate that glycosphingolipids expressing an appropriate oligosaccharide mediate cell tethering/rolling via E-selectin but not P-selectin.

Original language	English (US)
Pages (from-to)	42-49
Number of pages	8
Journal	Biochemical and Biophysical Research Communications
Volume	284
Issue number	1
DOIs	https://doi.org/10.1006/bbrc.2001.4899
State	Published - 2001

Funding

This work was supported by a Whitaker Foundation Grant, National Institutes of Health Grant AI45115, National Science Foundation Grant BES 9978160, and a National Science Foundation Graduate Research Fellowship (to M.M.B). 1Current address: The Scripps Research Institute, La Jolla, CA. 2To whom correspondence should be addressed at Department of Chemical Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218-2694. Fax: (410) 516-5510. E-mail: [email protected].

Keywords

Selectins
Shear
Sialyl Lewis

ASJC Scopus subject areas

Molecular Biology
Biophysics
Biochemistry
Cell Biology

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10.1006/bbrc.2001.4899

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title = "Glycolipids support E-selectin-specific strong cell tethering under flow",

abstract = "This study provides functional evidence that glycosphingolipids constitute ligands for E-selectin but not P-selectin. Chinese hamster ovary (CHO) cells expressing E-selectin (CHO-E) or P-selectin (CHO-P) were perfused over α2,3-sialyl Lewis X (α2,3-sLex) presented as the hexaosylceramide glycosphingolipid adsorbed in a monolayer containing phosphatidylcholine and cholesterol. CHO-E cells tethered extensively and formed slow, stable rolling interactions with α2,3-sLex glycosphingolipid but not with the comparable α2,6-sLex glycosphingolipid. Tethering/rolling varied with wall shear stress, selectin density, and ligand density. In contrast, α2,3-sLex glycosphingolipid supported only limited, fast CHO-P cell rolling. As calculated from a stochastic model of cell rolling, the step size between successive bond releases from the α2,3-sLex glycosphingolipid was smaller for CHO-E than CHO-P cells, whereas the opposite effect was observed for the waiting time between these events. Detachment assays revealed stronger adhesive interactions of CHO-E than CHO-P cells with α2,3-sLex glycosphingolipid. These findings indicate that glycosphingolipids expressing an appropriate oligosaccharide mediate cell tethering/rolling via E-selectin but not P-selectin.",

keywords = "Selectins, Shear, Sialyl Lewis",

author = "Burdick, {Monica M.} and Bochner, {Bruce S.} and Collins, {Brian E.} and Schnaar, {Ronald L.} and Konstantinos Konstantopoulos",

note = "Funding Information: This work was supported by a Whitaker Foundation Grant, National Institutes of Health Grant AI45115, National Science Foundation Grant BES 9978160, and a National Science Foundation Graduate Research Fellowship (to M.M.B). 1Current address: The Scripps Research Institute, La Jolla, CA. 2To whom correspondence should be addressed at Department of Chemical Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218-2694. Fax: (410) 516-5510. E-mail: [email protected].",

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AU - Konstantopoulos, Konstantinos

N1 - Funding Information: This work was supported by a Whitaker Foundation Grant, National Institutes of Health Grant AI45115, National Science Foundation Grant BES 9978160, and a National Science Foundation Graduate Research Fellowship (to M.M.B). 1Current address: The Scripps Research Institute, La Jolla, CA. 2To whom correspondence should be addressed at Department of Chemical Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218-2694. Fax: (410) 516-5510. E-mail: [email protected].

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N2 - This study provides functional evidence that glycosphingolipids constitute ligands for E-selectin but not P-selectin. Chinese hamster ovary (CHO) cells expressing E-selectin (CHO-E) or P-selectin (CHO-P) were perfused over α2,3-sialyl Lewis X (α2,3-sLex) presented as the hexaosylceramide glycosphingolipid adsorbed in a monolayer containing phosphatidylcholine and cholesterol. CHO-E cells tethered extensively and formed slow, stable rolling interactions with α2,3-sLex glycosphingolipid but not with the comparable α2,6-sLex glycosphingolipid. Tethering/rolling varied with wall shear stress, selectin density, and ligand density. In contrast, α2,3-sLex glycosphingolipid supported only limited, fast CHO-P cell rolling. As calculated from a stochastic model of cell rolling, the step size between successive bond releases from the α2,3-sLex glycosphingolipid was smaller for CHO-E than CHO-P cells, whereas the opposite effect was observed for the waiting time between these events. Detachment assays revealed stronger adhesive interactions of CHO-E than CHO-P cells with α2,3-sLex glycosphingolipid. These findings indicate that glycosphingolipids expressing an appropriate oligosaccharide mediate cell tethering/rolling via E-selectin but not P-selectin.

AB - This study provides functional evidence that glycosphingolipids constitute ligands for E-selectin but not P-selectin. Chinese hamster ovary (CHO) cells expressing E-selectin (CHO-E) or P-selectin (CHO-P) were perfused over α2,3-sialyl Lewis X (α2,3-sLex) presented as the hexaosylceramide glycosphingolipid adsorbed in a monolayer containing phosphatidylcholine and cholesterol. CHO-E cells tethered extensively and formed slow, stable rolling interactions with α2,3-sLex glycosphingolipid but not with the comparable α2,6-sLex glycosphingolipid. Tethering/rolling varied with wall shear stress, selectin density, and ligand density. In contrast, α2,3-sLex glycosphingolipid supported only limited, fast CHO-P cell rolling. As calculated from a stochastic model of cell rolling, the step size between successive bond releases from the α2,3-sLex glycosphingolipid was smaller for CHO-E than CHO-P cells, whereas the opposite effect was observed for the waiting time between these events. Detachment assays revealed stronger adhesive interactions of CHO-E than CHO-P cells with α2,3-sLex glycosphingolipid. These findings indicate that glycosphingolipids expressing an appropriate oligosaccharide mediate cell tethering/rolling via E-selectin but not P-selectin.

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Glycolipids support E-selectin-specific strong cell tethering under flow

Abstract

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Keywords

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