Interaction of tubulin with bifunctional colchicine analogues

An equilibrium study

Jose M. Andreu, Marina J. Gorbunoff, James Lee, Serge N. Timasheff

Research output: Contribution to journalArticle

106 Citations (Scopus)

Abstract

The interaction of tubulin with simple analogues of colchicine that contain both its tropolone and trimethoxyphenyl rings has been characterized, and the results were analyzed in terms of the simple bifunctional ligand model developed for the binding of colchicine [Andreu, J. M., & Timasheff, S. N. (1982) Biochemistry 21, 534-543] on the basis of interactions of tubulin with single-ring analogues. The compound 2-methoxy-5-(2,3,4-trimethoxyphenyl)-2,4,6-cycloheptatrien-1-one has been found to bind reversibly to 0.86 ± 0.06 site of purified calf brain tubulin with an equilibrium constant of (4.9 ± 0.3) × 105 M-1 (25 °C), ΔH°app = -1.6 ± 0.7 kcal mol-1, and ΔS°app = 20.5 ± 2.5 eu. The binding appears specific for the colchicine site. The closely related compound 2-methoxy-5-[[3-(3,4,5-trimethoxyphenyl)-propionyl]amino]-2,4,6-cycloheptatrien- 1-one interacts weakly with tubulin. Binding of the first analogue is accompanied by ligand fluorescence appearance, quenching of protein fluorescence, perturbation of the far-ultraviolet circular dichroism of tubulin, and induction of the tubulin GTPase activity, similarly to colchicine binding. Substoichiometric concentrations of the analogue inhibit microtubule assembly in vitro. Excess analogue concentration under microtubulepromoting conditions induces an abnormal cooperative polymerization of tubulin, similar to that of the tubulin-colchicine complex.

Original languageEnglish (US)
Pages (from-to)1742-1752
Number of pages11
JournalBiochemistry
Volume23
Issue number8
StatePublished - 1984
Externally publishedYes

Fingerprint

Colchicine
Tubulin
Tropolone
Fluorescence
Ligands
Biochemistry
GTP Phosphohydrolases
Equilibrium constants
Circular Dichroism
Microtubules
Polymerization
Quenching
Brain

ASJC Scopus subject areas

  • Biochemistry

Cite this

Andreu, J. M., Gorbunoff, M. J., Lee, J., & Timasheff, S. N. (1984). Interaction of tubulin with bifunctional colchicine analogues: An equilibrium study. Biochemistry, 23(8), 1742-1752.

Interaction of tubulin with bifunctional colchicine analogues : An equilibrium study. / Andreu, Jose M.; Gorbunoff, Marina J.; Lee, James; Timasheff, Serge N.

In: Biochemistry, Vol. 23, No. 8, 1984, p. 1742-1752.

Research output: Contribution to journalArticle

Andreu, JM, Gorbunoff, MJ, Lee, J & Timasheff, SN 1984, 'Interaction of tubulin with bifunctional colchicine analogues: An equilibrium study', Biochemistry, vol. 23, no. 8, pp. 1742-1752.
Andreu JM, Gorbunoff MJ, Lee J, Timasheff SN. Interaction of tubulin with bifunctional colchicine analogues: An equilibrium study. Biochemistry. 1984;23(8):1742-1752.
Andreu, Jose M. ; Gorbunoff, Marina J. ; Lee, James ; Timasheff, Serge N. / Interaction of tubulin with bifunctional colchicine analogues : An equilibrium study. In: Biochemistry. 1984 ; Vol. 23, No. 8. pp. 1742-1752.
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