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Fluorescence correlation spectroscopy analysis of the dynamics of tubulin interaction with RB3, a stathmin family protein
Author(s) -
Krouglova Tatiana,
Amayed Phedra,
Engelborghs Yves,
Carlier Marie-France
Publication year - 2003
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)00636-7
Subject(s) - stathmin , chemistry , tubulin , dissociation constant , dissociation (chemistry) , fluorescence correlation spectroscopy , kinetics , fluorescence , fluorescence spectroscopy , reaction rate constant , biophysics , stereochemistry , biochemistry , biology , microtubule , phosphorylation , molecule , organic chemistry , genetics , physics , receptor , quantum mechanics
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.