Solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates
The binding parameters, the affinity constant (K a ) and binding capacity (Q), of a protein possessing ligand-protein complexes with a high dissociation rate (Sex Steroid Binding protein from Bufo arenarum) were determined using a solid-phase method. The technique is based upon the adsorption of the...
Guardado en:
| Autores principales: | , , |
|---|---|
| Formato: | JOUR |
| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_00032697_v192_n2_p367_SantaColoma |
| Aporte de: |
| id |
todo:paper_00032697_v192_n2_p367_SantaColoma |
|---|---|
| record_format |
dspace |
| spelling |
todo:paper_00032697_v192_n2_p367_SantaColoma2023-10-03T13:55:48Z Solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates Santa-Coloma, T.A. Rossi, R.J. Charreau, E.H. sex hormone binding protein animal tissue article binding kinetics dissociation female ligand binding nonhuman priority journal protein binding Animal Bufo arenarum Carrier Proteins Charcoal Chromatography, DEAE-Cellulose Female Half-Life Kinetics Ligands Sex Hormone-Binding Globulin Sex Hormones Animalia Bufo arenarum The binding parameters, the affinity constant (K a ) and binding capacity (Q), of a protein possessing ligand-protein complexes with a high dissociation rate (Sex Steroid Binding protein from Bufo arenarum) were determined using a solid-phase method. The technique is based upon the adsorption of the steroid-protein complex to DEAE-cellulose. This method was compared with a nonequilibrium method (charcoal adsorption of free ligand), and the latter resulted in underestimation of both binding parameters, K a and Q. The solid-phase method reported here is appropriate to determine the binding parameters of proteins with high dissociation rates because the results are independent of the complex half-time. The method also possesses advantages compared to other equilibrium assays such as dialysis or steady-state electrophoresis. With minor modifications, it may be useful to characterize different proteins, particularly those possessing ligand-protein complexes with very high dissociation rates. © 1991. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00032697_v192_n2_p367_SantaColoma |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
sex hormone binding protein animal tissue article binding kinetics dissociation female ligand binding nonhuman priority journal protein binding Animal Bufo arenarum Carrier Proteins Charcoal Chromatography, DEAE-Cellulose Female Half-Life Kinetics Ligands Sex Hormone-Binding Globulin Sex Hormones Animalia Bufo arenarum |
| spellingShingle |
sex hormone binding protein animal tissue article binding kinetics dissociation female ligand binding nonhuman priority journal protein binding Animal Bufo arenarum Carrier Proteins Charcoal Chromatography, DEAE-Cellulose Female Half-Life Kinetics Ligands Sex Hormone-Binding Globulin Sex Hormones Animalia Bufo arenarum Santa-Coloma, T.A. Rossi, R.J. Charreau, E.H. Solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates |
| topic_facet |
sex hormone binding protein animal tissue article binding kinetics dissociation female ligand binding nonhuman priority journal protein binding Animal Bufo arenarum Carrier Proteins Charcoal Chromatography, DEAE-Cellulose Female Half-Life Kinetics Ligands Sex Hormone-Binding Globulin Sex Hormones Animalia Bufo arenarum |
| description |
The binding parameters, the affinity constant (K a ) and binding capacity (Q), of a protein possessing ligand-protein complexes with a high dissociation rate (Sex Steroid Binding protein from Bufo arenarum) were determined using a solid-phase method. The technique is based upon the adsorption of the steroid-protein complex to DEAE-cellulose. This method was compared with a nonequilibrium method (charcoal adsorption of free ligand), and the latter resulted in underestimation of both binding parameters, K a and Q. The solid-phase method reported here is appropriate to determine the binding parameters of proteins with high dissociation rates because the results are independent of the complex half-time. The method also possesses advantages compared to other equilibrium assays such as dialysis or steady-state electrophoresis. With minor modifications, it may be useful to characterize different proteins, particularly those possessing ligand-protein complexes with very high dissociation rates. © 1991. |
| format |
JOUR |
| author |
Santa-Coloma, T.A. Rossi, R.J. Charreau, E.H. |
| author_facet |
Santa-Coloma, T.A. Rossi, R.J. Charreau, E.H. |
| author_sort |
Santa-Coloma, T.A. |
| title |
Solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates |
| title_short |
Solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates |
| title_full |
Solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates |
| title_fullStr |
Solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates |
| title_full_unstemmed |
Solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates |
| title_sort |
solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates |
| url |
http://hdl.handle.net/20.500.12110/paper_00032697_v192_n2_p367_SantaColoma |
| work_keys_str_mv |
AT santacolomata solidphaseassayfordeterminationofbindingparametersofligandproteincomplexeswithhighdissociationrates AT rossirj solidphaseassayfordeterminationofbindingparametersofligandproteincomplexeswithhighdissociationrates AT charreaueh solidphaseassayfordeterminationofbindingparametersofligandproteincomplexeswithhighdissociationrates |
| _version_ |
1807321714227412992 |