Binding of anandamide to bovine serum albumin
The endocannabinoid anandamide is of lipid nature and may thus bind to albumin in the vascular system, as do fatty acids. The knowledge of the free water-phase concentration of anandamide is essential for the investigations of its transfer from the binding protein to cellular membranes, because a wa...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2003-09-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520337445 |
| _version_ | 1818444370392645632 |
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| author | Inge N. Bojesen Harald S. Hansen |
| author_facet | Inge N. Bojesen Harald S. Hansen |
| author_sort | Inge N. Bojesen |
| collection | DOAJ |
| description | The endocannabinoid anandamide is of lipid nature and may thus bind to albumin in the vascular system, as do fatty acids. The knowledge of the free water-phase concentration of anandamide is essential for the investigations of its transfer from the binding protein to cellular membranes, because a water-phase shuttle of monomers mediates such transfers. We have used our method based upon the use of albumin-filled red cell ghosts as a dispersed biological “reference binder” to measure the water-phase concentrations of anandamide. These concentrations were measured in buffer (pH 7.3) in equilibrium with anandamide bound to BSA inside resealed human red cell membranes at low molar ratios below one. Data were obtained at 0°C, 10°C, 23°C, and 37°C. The equilibrium dissociation constant (Kd) increases with temperature from 6.87 ± 0.53 nM at 0°C to 54.92 ± 1.91 nM at 37°C. Regression analyses of the data suggest that BSA has one high-affinity binding site for anandamide at all four temperatures. The free energy of anandamide binding (ΔG0) is calculated to −43.05 kJ mol−1 with a large enthalpy (ΔH0) contribution of −42.09 kJ mol−1.Anandamide has vasodilator activity, and the binding to albumin may mediate its transport in aqueous compartments. |
| first_indexed | 2024-12-14T19:14:51Z |
| format | Article |
| id | doaj.art-68192ec9e805490ca8886038d1205538 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-14T19:14:51Z |
| publishDate | 2003-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-68192ec9e805490ca8886038d12055382022-12-21T22:50:39ZengElsevierJournal of Lipid Research0022-22752003-09-0144917901794Binding of anandamide to bovine serum albuminInge N. Bojesen0Harald S. Hansen1Department of Medical Biochemistry and Genetics, Lab. B., University of Copenhagen, The Panum Institute, Blegdamsvej 3, DK-2200 Copenhagen N, Denmark; Department of Pharmacology, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen Ø, DenmarkDepartment of Medical Biochemistry and Genetics, Lab. B., University of Copenhagen, The Panum Institute, Blegdamsvej 3, DK-2200 Copenhagen N, Denmark; Department of Pharmacology, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen Ø, DenmarkThe endocannabinoid anandamide is of lipid nature and may thus bind to albumin in the vascular system, as do fatty acids. The knowledge of the free water-phase concentration of anandamide is essential for the investigations of its transfer from the binding protein to cellular membranes, because a water-phase shuttle of monomers mediates such transfers. We have used our method based upon the use of albumin-filled red cell ghosts as a dispersed biological “reference binder” to measure the water-phase concentrations of anandamide. These concentrations were measured in buffer (pH 7.3) in equilibrium with anandamide bound to BSA inside resealed human red cell membranes at low molar ratios below one. Data were obtained at 0°C, 10°C, 23°C, and 37°C. The equilibrium dissociation constant (Kd) increases with temperature from 6.87 ± 0.53 nM at 0°C to 54.92 ± 1.91 nM at 37°C. Regression analyses of the data suggest that BSA has one high-affinity binding site for anandamide at all four temperatures. The free energy of anandamide binding (ΔG0) is calculated to −43.05 kJ mol−1 with a large enthalpy (ΔH0) contribution of −42.09 kJ mol−1.Anandamide has vasodilator activity, and the binding to albumin may mediate its transport in aqueous compartments.http://www.sciencedirect.com/science/article/pii/S0022227520337445equilibrium dissociation constantresealed red cell membraneserythrocyte ghostsequilibrium constant of anandamide-albumin complexanandamide monomer concentration |
| spellingShingle | Inge N. Bojesen Harald S. Hansen Binding of anandamide to bovine serum albumin Journal of Lipid Research equilibrium dissociation constant resealed red cell membranes erythrocyte ghosts equilibrium constant of anandamide-albumin complex anandamide monomer concentration |
| title | Binding of anandamide to bovine serum albumin |
| title_full | Binding of anandamide to bovine serum albumin |
| title_fullStr | Binding of anandamide to bovine serum albumin |
| title_full_unstemmed | Binding of anandamide to bovine serum albumin |
| title_short | Binding of anandamide to bovine serum albumin |
| title_sort | binding of anandamide to bovine serum albumin |
| topic | equilibrium dissociation constant resealed red cell membranes erythrocyte ghosts equilibrium constant of anandamide-albumin complex anandamide monomer concentration |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520337445 |
| work_keys_str_mv | AT ingenbojesen bindingofanandamidetobovineserumalbumin AT haraldshansen bindingofanandamidetobovineserumalbumin |