Summary
11-cis retinaldehyde binding analysis was performed on a bovine retinal pigment epithelium preparation of cellular retinaldehyde binding protein (CRALBP), whose purity degree was estimated as 75%. Equilibrium binding studies were carried out measuring the replacement of tritium-labeled with unlabeled 11-cis retinaldehyde at 25°C. Analysis of the experimental data both by a direct curve-fitting procedure utilizing a non linear least square regression analysis and by a conventional Scatchard plot revealed a single non-interacting binding site with an apparent equilibrium constant of 0.9×10−7 M.
A binding stoichiometry of approximately 1 mol of 11-cis retinaldehyde/mol of binding protein can be calculated from the experimental data. Competition studies carried out in the presence of unlabeled ‘trans’ and ‘cis’ isomers of Vitamin A derivatives confirm the high degree of specificity of the 11-cis retinaldehyde binding.
References
Futterman, S. Saari, J.C., and Blair, S., J. biol. Chem.252 (1977) 3267.
Stubbs, G.W., Saari, J.C., and Futterman, S., J. biol. Chem.254 (1979) 8529.
Saari, J.C., Bredberg, L., and Garwin, G.G., J. biol. Chem.257 (1982) 13329.
Saari, J.C., Teller, D.C., Crabb, J.W., and Bredberg, L., J. biol. Chem.260 (1985) 195.
Bunt-Milam, A.H., and Saari, J.C., J. Cell Biol.97 (1983) 703.
Rothmans, J.P., and Kropf, A., Vision Res.15 (1975) 1301.
Groenendijk, G.W.T., Jacobs, C.W.M., Bonting, S.L., and Daemen, F.J.M., Eur. J. Biochem.106 (1980) 119.
Groenendijk, G.W.T., Jansen, P.A.A., Bonting, S.L., and Daemen, F.J.M., in: Methods in Enzymology, vol. 67, p. 203. Eds D.B. Mc-Cormick and L.D. Wright. Academic Press, New York 1980.
Feeney-Burns, L., and Berman, E., in: Methods in Enzymology, vol. 81, p. 95. Ed. L. Packer. Academic Press, New York 1982.
Saari, J.C., in: Methods in Enzymology, vol. 81, p. 819. Ed. L. Packer. Academic Press, New York 1982.
Zahler, W.L., in: Methods in Enyzmology, vol. 32, part B, p. 70. Eds S. Fleischer and L. Packer. Academic Press, New York 1974.
Saari, J.C., Bunt-Milam, A.H., Bredberg, D.L., and Garwin, G.G., Vision Res.24 (1984) 1595.
Bashor, M.M., Toft, D.O., and Chytil, F., Proc. natn. Acad. Sci. USA70 (1973) 3483.
Bashor, M.M., and Chytil, F., Biochim. biophys. acta411 (1975) 87.
Bradford, M.M., Analyt. Biochem.72 (1976) 248.
Atkins, G.L., Eur. J. Biochem.33 (1973) 175.
Bridges, C.D.B., Alvarez, R.A., Fong, S.L., Gonzalez-Fernandez, F., Lam, M.D.K., and Liou, G.I., Vision Res.24 (1984) 1581.
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Livrea, M.A., Bongiorno, A., Tesoriere, L. et al. Binding of 11-cis retinaldehyde to the partially purified cellular retinaldehyde binding protein from bovine retinal pigment epithelium. Experientia 43, 582–586 (1987). https://doi.org/10.1007/BF02143594
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DOI: https://doi.org/10.1007/BF02143594