Isolation of α and β brain tubulin subunits after alkaline treatment of the protein

Dante M. Beltramo, Mariana Nuñez Fernandez, Alejandra Del C. Alonso, Juan J. Sironi, Héctor S. Barra

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2 Scopus citations

Abstract

We demonstrate here that brain purified tubulin can be dissociated into α and β subunits at pH > 10 and that the subunits can be separated by using the Triton X-114 phase separation system. After phase partition at pH > 10, α tubulin but not β tubulin behaves as a hydrophobic compound appearing in the detergent rich phase. After three extractions of the alkaline aqueous phase with Triton X-114, about 90% of the α tubulin was recovered in the detergent rich phase. The hydrophobic behavior observed for α tubulin after its dissociation at pH 11.5 was not due to an irreversible change of the protein, because when the detergent rich phase containing α tubulin was diluted with a buffer solution at pH 7.3 and the solution allowed to partition again, α-tubulin is recovered in the aqueous phase. The detergent in the aqueous phase of the α and β tubulin preparations can be removed up to 90% by 12 h dialysis. The α and β subunits of tubulin from kidney and liver behave, in this phase separation system, like those of brain tubulin.

Original languageEnglish (US)
Pages (from-to)385-389
Number of pages5
JournalNeurochemical Research
Volume22
Issue number4
DOIs
Publication statusPublished - Jan 1 1997

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Keywords

  • Brain
  • alpha tubulin
  • beta tubulin
  • tubulin
  • tubulin subunits separation

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Beltramo, D. M., Fernandez, M. N., Del C. Alonso, A., Sironi, J. J., & Barra, H. S. (1997). Isolation of α and β brain tubulin subunits after alkaline treatment of the protein. Neurochemical Research, 22(4), 385-389. https://doi.org/10.1023/A:1027395307699