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Abstract:

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.

Registro:

Documento: Artículo
Título:Solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates
Autor:Santa-Coloma, T.A.; Rossi, R.J.; Charreau, E.H.
Filiación:Instituto de Biología y Medicina Experimental, Obligado 2490, Buenos Aires, 1428, Argentina
Palabras clave: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
Año:1991
Volumen:192
Número:2
Página de inicio:367
Página de fin:371
DOI: http://dx.doi.org/10.1016/0003-2697(91)90550-D
Título revista:Analytical Biochemistry
Título revista abreviado:Anal. Biochem.
ISSN:00032697
CODEN:ANBCA
CAS:Carrier Proteins; Charcoal, 16291-96-6; Ligands; Sex Hormone-Binding Globulin; Sex Hormones
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032697_v192_n2_p367_SantaColoma

Referencias:

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Citas:

---------- APA ----------
Santa-Coloma, T.A., Rossi, R.J. & Charreau, E.H. (1991) . Solid-phase assay for determination of binding parameters of ligand-protein complexes with high dissociation rates. Analytical Biochemistry, 192(2), 367-371.
http://dx.doi.org/10.1016/0003-2697(91)90550-D
---------- CHICAGO ----------
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" . Analytical Biochemistry 192, no. 2 (1991) : 367-371.
http://dx.doi.org/10.1016/0003-2697(91)90550-D
---------- MLA ----------
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" . Analytical Biochemistry, vol. 192, no. 2, 1991, pp. 367-371.
http://dx.doi.org/10.1016/0003-2697(91)90550-D
---------- VANCOUVER ----------
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. Anal. Biochem. 1991;192(2):367-371.
http://dx.doi.org/10.1016/0003-2697(91)90550-D