Affinity, kinetics, and thermodynamics of E-selectin binding to E-selectin ligand-1.
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...
| Huvudupphovsmän: | , , , , |
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| Materialtyp: | Journal article |
| Språk: | English |
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2001
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| _version_ | 1826292113168400384 |
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| author | Wild, M Huang, M Schulze-Horsel, U Van Der Merwe, P Vestweber, D |
| author_facet | Wild, M Huang, M Schulze-Horsel, U Van Der Merwe, P Vestweber, D |
| author_sort | Wild, M |
| collection | OXFORD |
| description | 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 x 10(4) m(-1) s(-1). We determined a dissociation constant (K(d)) of 62 microm, which resembles the affinity of L-selectin binding to glycosylation-dependent cell adhesion molecule-1. The affinity of the E-selectin-ESL-1 interaction did not change significantly when the temperature was varied from 5 degrees C to 37 degrees 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 alpha 1,3-fucosyltransferase IV-expressing Chinese hamster ovary cells showed a very similar K(d) of 66 microm, 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. |
| first_indexed | 2024-03-07T03:09:41Z |
| format | Journal article |
| id | oxford-uuid:b3c2064b-097a-4cbd-bfa2-c9370a22f9f1 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:09:41Z |
| publishDate | 2001 |
| record_format | dspace |
| spelling | oxford-uuid:b3c2064b-097a-4cbd-bfa2-c9370a22f9f12022-03-27T04:21:30ZAffinity, kinetics, and thermodynamics of E-selectin binding to E-selectin ligand-1.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:b3c2064b-097a-4cbd-bfa2-c9370a22f9f1EnglishSymplectic Elements at Oxford2001Wild, MHuang, MSchulze-Horsel, UVan Der Merwe, PVestweber, DE-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 x 10(4) m(-1) s(-1). We determined a dissociation constant (K(d)) of 62 microm, which resembles the affinity of L-selectin binding to glycosylation-dependent cell adhesion molecule-1. The affinity of the E-selectin-ESL-1 interaction did not change significantly when the temperature was varied from 5 degrees C to 37 degrees 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 alpha 1,3-fucosyltransferase IV-expressing Chinese hamster ovary cells showed a very similar K(d) of 66 microm, 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. |
| spellingShingle | Wild, M Huang, M Schulze-Horsel, U Van Der Merwe, P Vestweber, D Affinity, kinetics, and thermodynamics of E-selectin binding to E-selectin ligand-1. |
| title | Affinity, kinetics, and thermodynamics of E-selectin binding to E-selectin ligand-1. |
| title_full | Affinity, kinetics, and thermodynamics of E-selectin binding to E-selectin ligand-1. |
| title_fullStr | Affinity, kinetics, and thermodynamics of E-selectin binding to E-selectin ligand-1. |
| title_full_unstemmed | Affinity, kinetics, and thermodynamics of E-selectin binding to E-selectin ligand-1. |
| title_short | Affinity, kinetics, and thermodynamics of E-selectin binding to E-selectin ligand-1. |
| title_sort | affinity kinetics and thermodynamics of e selectin binding to e selectin ligand 1 |
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