Abstract
At least two types of cytokinin-binding sites are present in a particulate fraction of tobacco (Nicotiana tabacum L.) cells that sediments at 80,000 x g. The major binding component has a low affinity towards cytokinins, is resistant to heating at 100°C, and is not specific for biologically active cytokinin analogues. The second site occurs in much lower frequency, is heat labile, shows high affinity towards cytokinins, and is specific for biologically active analogs of the hormone. The testing for binding specificity was mainly performed with a series of halogenated benzyladenine derivatives having a wide range of biological activities. The low-affinity binding site shows some of the same features as talcum powder, a non-biological material which binds cytokinins in a non-specific fashion. The properties of the high-affinity binding site are consistent with the expected characteristics of a cytokinin receptor. However, the role of the observed high-affinity binding site with regard to the biological action of cytokinins is not yet known.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- BA:
-
N 6-benzyladenine
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- IAA:
-
indole-3-acetic acid
- Kd :
-
equilibrium dissociation constant
- Rt :
-
total concentration of binding sites
References
Andreev, G.K., Amrhein, N.: Mechanism of action of the herbicide 2-chloro-3(4-chlorophenyl)propionate and its methyl ester: interactions with cell responses mediated by auxin. Physiol. Plant. 37, 175–182 (1976)
Cuatrecasas, P., Hollenberg, M.D.: Binding of insulin and other hormones to non-receptor materials: saturability, specificity and apparent “negative cooperativity”. Biochem. Biophys. Res. Commun. 62, 31–41 (1975)
Cuatrecasas, P., Hollenberg, M.D.: Membrane receptors and hormone action. Adv. Protein Chem. 30, 251–451 (1976)
Cuatrecasas, P., Hollenberg, M.D., Chang, K., Bennett, V.: Hormone receptor complexes and their modulation of membrane function. Recent Progr. Hormone Res. 31, 37–94 (1975)
Cuatrecasas, P., Tell, C.P.E., Sica, V., Parikh, I., Chang, K.: Noiadrenalin binding and the search for catecholamine receptors. Nature 241, 92–97 (1974)
Elion, G.B., Burgi, E., Hitchings, G.H.: Studies on condensed pyrimidine systems. IX. The synthesis of some 6-substituted purines. J. Amer. Chem. Soc. 74, 411–414 (1952)
Fox, J.E., Erion, J.L.: A cytokinin binding protein from higher plant ribosomes. Biochem. Biophys. Res. Commun. 64, 694–700 (1975)
Hertel, R., Thomson, K.St., Russo, V.E.A.: In-vitro auxin binding to particulate cell fractions from corn coleoptiles. Planta 107, 325–340 (1972)
Jouanneau, J.P., Péaud-Lenoöl, C.: Croissance et synthèse des protéines de suspensions cellulaires de Tabac sensibles à la kinétine. Physiol. Plant. 20, 834–850 (1967)
Kende, H., Gardner, G.: Hormone binding in plants. Ann. Rev. Plant Physiol. 27, 267–290 (1976)
Kende, H., Hahn, H., Kays, S.E.: Enhancement of nitrate reductase by benzyladenine in Agrostemma githago. Plant Physiol. 48, 702–706 (1971)
Kuraishi, S.: Effect of kinetin analogs on leaf growth. Sci. Paps., Coll. Gen. Educ., Univ. Tokyo 9, 67–104 (1959)
Linsmaier, E.M., Skoog, F.: Organic growth factor requirements of tobacco tissue cultures. Physiol. Plant. 18, 100–127 (1965)
Okumura, F.S., Kotani, Y., Ariga, T., Masumura, M., Kuraishi, S.: Syntheses of kinetin-analogues. Bull. Chem. Soc. Japan 32, 883–887 (1959)
Puca, G.A., Nola, E., Sica, V., Bresciani, F.: Estrogen-binding proteins of calf uterus. Partial purification and preliminary characterization of two cytoplasmic proteins. Biochemistry 10, 3769–3779 (1971)
Ray, P.M., Dohrmann, U., Hertel, R.: Characterization of naphthalene-acetic acid binding to receptor sites on cellular membranes of maize coleoptile tissue. Plant Physiol. 59, 357–364 (1977)
Rosenthal, H.E.: A graphic method for the determination and presentation of binding parameters in a complex system. Anal. Biochem. 20, 525–532 (1967)
Scatchard, G.: The attractions of proteins for small molecules and ions. Ann. N.Y. Acad. Sci. 51, 660–672 (1949)
Sussman, M.R.: Studies on cytokinin binding in tobacco cells. Ph.D. thesis, Michigan State University, East Lansing, Mich., USA 1976. Diss. Abstr. 38, 1028-B (1977)
Sussman, M.R., Firn, R.D.: The synthesis of a radioactive cytokinin with high specific activity. Phytochem. 15, 153–155 (1976)
Sussman, M.R., Kende, H.: The synthesis and biological properties of 8-azido-N 6-benzyladenine, a potential photoaffinity reagent for cytokinin. Planta 137, 91–96 (1977)
Takegami, T., Yoshida, K.: Isolation and purification of cytokinin binding protein from tobacco leaves by affinity chromatography. Biochem. Biophys. Res. Commun. 67, 782–789 (1975)
Tandeau de Marsac, N., Jouanneau, J.: Variation de l'exigence en cytokinine de lignées clonales de cellules de Tabac. Physiol. vég. 10, 369–380 (1972)
Theiler, J.B., Leonard, N.J., Schmitz, R.Y., Skoog, F.: Photoaffinity-labeled cytokinins. Plant Physiol. 58, 803–805 (1976)
Whitaker, B.D., Kende, H.: Bud formation in Funaria hygrometrica: a comparison of the activities of three cytokins with their ribosides. Planta 121, 93–96 (1974)
Author information
Authors and Affiliations
Additional information
In partial fulfillment of the requirements for the Ph.D. degree in the Department of Botany and Plant Pathology, Michigan State University
Rights and permissions
About this article
Cite this article
Sussman, M.R., Kende, H. In vitro cytokinin binding to a particulate fraction of tobacco cells. Planta 140, 251–259 (1978). https://doi.org/10.1007/BF00390256
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00390256