Heat-shock protein 90 (hsp90) binds in vitro to tubulin dimer and inhibits microtubule formation.
Hsp90 interacts with steroid hormone receptors, protein kinases, and cytoskeletal proteins. The mode of action of hsp90 on microtubules and tubulin has not been investigated. Using isolated purified hsp90 and isolated tubulin, we demonstrated in vitro by difference absorption and fluorescence spectroscopy that hsp90 bound to tubulin with an apparent affinity constant of 5 x 10(5) M-1, assuming an apparent stoichiometry of 1 at 25 degrees C. Using microcalorimetry, we found a delta H of -9.8 +/- 0.8 kJ.mol-1. The binding of hsp90 to tubulin was confirmed by a sedimentation assay. Moreover, we showed that hsp90 inhibited tubulin polymerisation.
Garnier
C
C
Faculté de Pharmacie, UPRESA CNRS 6032, Marseille, France.
Barbier
P
P
Gilli
R
R
Lopez
C
C
Peyrot
V
V
Briand
C
C
D000818:Animals; D019281:Dimerization; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D013552:Swine; D014404:Tubulin / Q000378:metabolism*
D018841:HSP90 Heat-Shock Proteins; D014404:Tubulin