Binding of Sialyl Lewis X to E-Selectin As Measured by Fluorescence Polarization

Jacob, Gary S.; Kirmaier, Christine; Abbas, Saeed A.; Howard, Susan C.; Steininger, Christina N.; Welply, Joseph K.; Scudder, Peter

doi:10.1021/bi00004a014

Cited by 85 publications

(53 citation statements)

References 35 publications

(47 reference statements)

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“…Three of the studies avoided this problem by studying direct binding of sLe x to E-selectin (63,64,66). Still, these studies reported rather different K d -values of 107, 720, and 1100 -1800 M, respectively.…”

Section: Table II Summary Of Affinity Measurements Of E-selectin Bindmentioning

confidence: 99%

Affinity, Kinetics, and Thermodynamics of E-selectin Binding to E-selectin Ligand-1

Wild¹,

Huang²,

Schulze-Horsel³

et al. 2001

Journal of Biological Chemistry

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E-selectin is an endothelial adhesion molecule, which mediates the tethering and rolling of leukocytes on vascular endothelium. It recognizes the glycoprotein E-selectin ligand-1 (ESL-1) as a major binding partner on mouse myeloid cells. Using surface plasmon resonance, we measured the kinetics and affinity of binding of monomeric E-selectin to ESL-1 isolated from mouse bone marrow cells. E-selectin bound to ESL-1 with a fast dissociation rate constant of 4.6 s ؊1 and a calculated association rate constant of 7.4 ؋ 10 4 M ؊1 s ؊1 . We determined a dissociation constant (K d ) of 62 M, which resembles the affinity of L-selectin binding to glycosylationdependent cell adhesion molecule-1. The affinity of the E-selectin-ESL-1 interaction did not change significantly when the temperature was varied from 5°C to 37°C, indicating that the enthalpic contribution to the binding is small at physiological temperatures, and that, in contrast to typical protein-carbohydrate interactions, binding is driven primarily by favorable entropic changes. Interestingly, surface plasmon resonance experiments with recombinant ESL-1 from ␣1,3-fucosyltransferase IV-expressing Chinese hamster ovary cells showed a very similar K d of 66 M, suggesting that this fucosyltransferase is sufficient to produce fully functional recombinant ESL-1. Following the recent description of the affinity and kinetics of the selectin-ligand pairs L-selectin-glycosylation-dependent cell adhesion molecule-1 and P-selectin-P-selectin glycoprotein ligand-1, this is the first determination of the parameters of E-selectin binding to one of its naturally occurring ligands.

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Section: Table II Summary Of Affinity Measurements Of E-selectin Bindmentioning

confidence: 99%

Affinity, Kinetics, and Thermodynamics of E-selectin Binding to E-selectin Ligand-1

Wild¹,

Huang²,

Schulze-Horsel³

et al. 2001

Journal of Biological Chemistry

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“…Selectins have been shown to bind synthetic oligosaccharides related to sialylated and/or sulfated Lewis x (Le x , galactose ␤134(fucose ␣133)(N-acetyl)glucosamine) or its stereoisomer Lewis a (Le a , galactose ␤133(fucose ␣134)(Nacetyl)glucosamine) with very low affinities (K d 0.1-5 mM (11)(12)(13)(14)(15)(16)(17)(18)(19)). It is possible that these studies underestimated the affinities because: (i) with few exceptions (13,18,19), they were based on inhibition by synthetic oligosaccharides of multivalent selectin-ligand interactions, and (ii) these oligosaccharides may differ in structure from physiological selectin ligands (7). Indeed soluble, recombinant forms of CD62P (20) and CD62E (21) have been reported to bind leukocytes with much higher affinities (K d Յ 1 M).…”

Section: ؊1mentioning

confidence: 99%

Affinity and Kinetic Analysis of L-selectin (CD62L) Binding to Glycosylation-dependent Cell-adhesion Molecule-1

Nicholson¹,

Barclay²,

Singer³

et al. 1998

Journal of Biological Chemistry

186

117

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The selectin family of cell adhesion molecules mediates the tethering and rolling of leukocytes on blood vessel endothelium. It has been postulated that the molecular basis of this highly dynamic adhesion is the low affinity and rapid kinetics of selectin interactions. However, affinity and kinetic analyses of monomeric selectins binding their natural ligands have not previously been reported. Leukocyte selectin (L-selectin, CD62L) binds preferentially to O-linked carbohydrates present on a small number of mucin-like glycoproteins, such as glycosylation-dependent cell adhesion molecule-1 (Gly-CAM-1), expressed in high endothelial venules. Gly-CAM-1 is a soluble secreted protein which, following binding to CD62L, stimulates ␤ 2 -integrin-mediated adhesion of lymphocytes. Using surface plasmon resonance, we show that a soluble monomeric form of CD62L binds to purified immobilized GlyCAM-1 with a dissociation constant (K d ) of 108 M. CD62L dissociates from GlyCAM-1 with a very fast dissociation rate constant (>10 s ؊1 ) which agrees well with the reported dissociation rate constant of CD62L-mediated leukocyte tethers. The calculated association rate constant is >10 M ؊1 s ؊1. At concentrations just above its mean serum level (ϳ1.5 g/ml or ϳ30 nM), GlyCAM-1 binds multivalently to immobilized CD62L. It follows that soluble GlyCAM-1 may cross-link CD62L when it binds to cells, suggesting a mechanism for signal transduction.

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“…The molecular recognition of these nanoparticles was investigated by their binding to D-galactose-specific lectin, RCA 120 17) . The binding affinities of galactosides were determined directly by competitive binding to the lectin with a fluorescent sugar, 4-methylumbelliferyl b-D-galactopyranoside (MUG) 18,19) .…”

Section: Introductionmentioning

confidence: 99%

Self-aggregate nanoparticles of cholesteryl and galactoside groups-substituted pullulan and their specific binding to galactose specific lectin, RCA120

Taniguchi

Akiyoshi

Sunamoto³

1999

Macromol. Chem. Phys.

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SUMMARY:The hydrophobized polysaccharide cholesterol-bearing pullulan (CHP) forms hydrogel nanoparticles upon self-aggregation in water. To endow cell specificity to nanoparticles of CHP, galactosides were additionally substituted to CHP with various spacer lengths and degrees of substitution. The resulting CHP derivatives also form monodisperse nanoparticles by self-aggregation in water. All self-aggregate nanoparticles strongly bind to the b-D-galactose-specific lectin, RCA 120 (Ricinus communis agglutinin). The affinity of substituted galactosides to the lectin was quantitatively studied using a fluorescence polarization technique by competitive binding with 4-methylumbelliferyl b-D-galactopyranoside (MUG). This affinity strongly depends on its chemical structure, degree of substitution, and spacer length. The binding constant per galactose moiety drastically increased by substitution to the polymer backbone.

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Binding of Sialyl Lewis X to E-Selectin As Measured by Fluorescence Polarization

Cited by 85 publications

References 35 publications

Affinity, Kinetics, and Thermodynamics of E-selectin Binding to E-selectin Ligand-1

Affinity, Kinetics, and Thermodynamics of E-selectin Binding to E-selectin Ligand-1

Affinity and Kinetic Analysis of L-selectin (CD62L) Binding to Glycosylation-dependent Cell-adhesion Molecule-1

Self-aggregate nanoparticles of cholesteryl and galactoside groups-substituted pullulan and their specific binding to galactose specific lectin, RCA120

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