Interaction of vinblastine with calf brain microtubule protein

J. C. Lee, D. Harrison, S. N. Timasheff

Research output: Contribution to journalArticle

133 Scopus citations

Abstract

The interaction of vinblastine with calf brain tubulin has been studied by velocity sedimentation, gel filtration, and fluorescence. It has been established that vinblastine induces the stable tubulin dimers to dimerize further to tetramers. The sedimentation patterns at low vinblastine concentration were analyzed by the ligand induced dimerization theory of Cann and Goad. The association constant and stoichiometry for the binding of vinblastine to tubulin, determined by gel filtration and spectrofluorometry, were (2.3±0.1) x 104 liters/mol at 25° and two vinblastine binding sites per tubulin dimer of molecular weight 110,000. The binding of vinblastine to tubulin is characterized by an enthalpy change of 5.8 kcal/mol and a positive unitary entropy change. Binding of vinblastine did not induce any significant conformational changes in tubulin as monitored by circular dichroism. However, the vinblastine tubulin complex displayed an ultraviolet different spectrum, which appears to reflect mostly the transfer of vinblastine tubulin complex displayed an ultraviolet difference spectrum, which appears to reflect mostly the transfer of vinblastine to a less polar environment. Besides binding vinblastine, tubulin was shown to bind vincristine with identical free energy and stoichiometry and to have a single binding site for 8 anilino 1 naphthalene sulfonic acid per tubulin dimer, which is independent of those for vinblastine.

Original languageEnglish (US)
Pages (from-to)9276-9282
Number of pages7
JournalJournal of Biological Chemistry
Volume250
Issue number24
StatePublished - Dec 1 1975

    Fingerprint

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lee, J. C., Harrison, D., & Timasheff, S. N. (1975). Interaction of vinblastine with calf brain microtubule protein. Journal of Biological Chemistry, 250(24), 9276-9282.