Conformational study of calf brain tubulin

James C. Lee, Debra Corfman, Ronald P. Frigon, Serge N. Timasheff

Research output: Contribution to journalArticle

41 Scopus citations

Abstract

The conformation of calf brain tubulin has been monitored by circular dichroism, optical rotatory dispersion, and spectrophotometric titration as a function of pH, temperature, ligand concentrations, and denaturants. At pH 7, calf brain tubulin maintains its structural integrity between 5 and 37 °C as determined by circular dichroism. Furthermore, the presence of MgCl2 up to 1.6 × 10-2m does not induce any observable changes in the circular dichroism spectra, nor does 10-4m CaCl2. With increasing pH, the spectral data can best be described as a gradual loosening of the secondary structure between pH 7 and 9. Both spectral and titrimetric data suggest a major unfolding of tubulin between pH 9 and 10. The apparent pK of tyrosine shifts from 10.85 to 9.98 upon transferring from buffer to 6 m guanidine hydrochloride, indicating that at least 14 of the 15 tyrosine groups are not fully accessible to protons in the native protein. The single disulfide bridge in calf brain tubulin helps to maintain a domain which is highly resistant to unfolding by denaturants.

Original languageEnglish (US)
Pages (from-to)4-14
Number of pages11
JournalArchives of Biochemistry and Biophysics
Volume185
Issue number1
DOIs
StatePublished - Jan 15 1978

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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    Lee, J. C., Corfman, D., Frigon, R. P., & Timasheff, S. N. (1978). Conformational study of calf brain tubulin. Archives of Biochemistry and Biophysics, 185(1), 4-14. https://doi.org/10.1016/0003-9861(78)90137-6