title: Thermodynamics of colchicinoid-tubulin interactions. Rrol of B-ring and C-7 substituent.
abstract: The quenching of tryptophan fluorescence has been used to determine the kinetic and thermodynamic parameters of binding of B-ring analogs of colchicine to tubulin. The on rate, activation energy, off-rate, and thermodynamics of binding reaction have been found to be controlled at different points of analog structure. The on-rate and off-rate of deacetamidocolchicine (DAAC) binding with tubulin is 17 times slower than that of 2-methoxy-5-(2',3',4'-trimethoxyphenyl)tropone-tubulin (AC-tubulin) interaction, although both reactions have very similar activation energies. The presence of B-ring alone does not significantly affect the thermodynamics of the binding reactions either, since both AC-tubulin and DAAC-tubulin interactions are enthalpy driven. Introduction of a NH2 group at C-7 position of the B-ring, as in deacetylcolchicine (NH2-DAAC) lowers the on-rate further with a significant rise in the value of the activation energy. However, bulkier substitutions at the same position, as in demecolcine (NHMe-DAAC) and N-methyldemecolcine (NMe2-DAAC) have no significant additional effect either on the on-rate or on the value of activation energy. Introduction of NH2 group in the C-7 position of B-ring also increases the positive entropy of the binding reaction to a significant extent, and it is maximum when NMe2 is substituted instead of NH2 group. Thus, interaction of NH2-DAAC, NHMe-DAAC, and NMe2-DAAC with tubulin are entropy driven. Our results suggest that the B-ring side chain of aminocolchicinoids makes contact(s) with dimeric tubulin molecules.
authors.affiliation: Department of Biochemistry, Bose Institute, Centenary Building, Calcutta 700 054, India.
authors.forename: G
authors.initials: G
authors.lastname: Chakrabarti
authors.forename: S
authors.initials: S
authors.lastname: Sengupta
authors.forename: B
authors.initials: B
authors.lastname: Bhattacharyya
mesh_terms: D000818:Animals; D001665:Binding Sites; D001921:Brain; D003078:Colchicine / Q000031:analogs & derivatives* / Q000737:chemistry*; D003703:Demecolcine / Q000737:chemistry; D006041:Goats; D007700:Kinetics; D008433:Mathematics; D008962:Models, Theoretical; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D014364:Tryptophan; D014404:Tubulin / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D014404:Tubulin; D014364:Tryptophan; D003078:Colchicine; D003703:Demecolcine