Fluorescence correlation spectroscopy analysis of the dynamics of tubulin interaction with RB3, a stathmin family protein

FEBS Letters

Amayed, Phedra¹  Carlier, Marie-France¹  Engelborghs, Yves²  Krouglova, Tatiana² 

[1] Dynamique du Cytosquelette, Laboratoire d'Enzymologie et Biochimie Structurales, CNRS, 91198 Gif-sur-Yvette, France;Laboratory of Biomolecular Dynamics, University of Leuven, Celestijnenlaan 200D, B-3001 Leuven, Belgium

关键词: Fluorescence correlation spectroscopy;    Tubulin;    RB3 protein;    Association–dissociation kinetics;    FCS;    fluorescence correlation spectroscopy;    TAMRA;    carboxytetramethylrhodamine succinimidyl ester;    T2R;    two tubulin dimers in complex with RB3;   

DOI  :  10.1016/S0014-5793(03)00636-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have used fluorescence correlation spectroscopy to analyze the interaction of GTP-tubulin with rhodamine-labeled RB3, a neural protein of the stathmin family, and to determine the kinetic pathway of the association process. RB3 displayed slow association–dissociation kinetics with tubulin depending on the square of the tubulin concentration. The values of the apparent association and dissociation rate constants of the complex of two tubulin dimers and RB3 are determined to be (3.52±0.14)×10⁻³ μM⁻²/s and (1.9±0.6)×10⁻³ s⁻¹ respectively. The value of the equilibrium dissociation constant for the first tubulin–RB3 interaction is estimated to be ⩾7 μM at 20°C.

【 授权许可】

Unknown