The chemical characterization of calf brain microtubule protein subunits

James Lee, R. P. Frigon, S. N. Timasheff

Research output: Contribution to journalArticle

187 Citations (Scopus)

Abstract

Calf brain microtubule protein (tubulin) was characterized chemically. Amino acid analysis and hydrogen ion titration in 5 M guanidine hydrochloride yielded a chemical composition similar to that of tubulin from other sources. The NH2 terminal residue was identified as methionine. When the protein was denatured and reduced in 8 M urea, two distinct protein bands were observed in gel electrophoresis in the presence of urea, suggesting the existence of two nonidentical subunits. The molecular weight of the tubulin subunits was determined by a variety of techniques, including sedimentation equilibrium and light scattering in 6 M guanidine hydrochloride, sodium dodecyl sulfate gel electrophoresis, chromatography on an agarose column in the presence of 6 M guanidine hydrochloride, and viscometry of S carboxymethylated, reduced protein in 6 M guanidine hydrochloride. All methods yielded an average molecular weight value of (54,000 ± 1,000), whether the measurements were done with or without reducing agents, indicating that all disulfide bonds present are intrachain.

Original languageEnglish (US)
Pages (from-to)7253-7262
Number of pages10
JournalJournal of Biological Chemistry
Volume248
Issue number20
StatePublished - 1973
Externally publishedYes

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Microtubule Proteins
Guanidine
Protein Subunits
Brain
Tubulin
Electrophoresis
Urea
Molecular Weight
Gels
Molecular weight
Proteins
Reducing Agents
Viscosity measurement
Chromatography
Titration
Sedimentation
Sodium Dodecyl Sulfate
Disulfides
Methionine
Light scattering

ASJC Scopus subject areas

  • Biochemistry

Cite this

Lee, J., Frigon, R. P., & Timasheff, S. N. (1973). The chemical characterization of calf brain microtubule protein subunits. Journal of Biological Chemistry, 248(20), 7253-7262.

The chemical characterization of calf brain microtubule protein subunits. / Lee, James; Frigon, R. P.; Timasheff, S. N.

In: Journal of Biological Chemistry, Vol. 248, No. 20, 1973, p. 7253-7262.

Research output: Contribution to journalArticle

Lee, J, Frigon, RP & Timasheff, SN 1973, 'The chemical characterization of calf brain microtubule protein subunits', Journal of Biological Chemistry, vol. 248, no. 20, pp. 7253-7262.
Lee, James ; Frigon, R. P. ; Timasheff, S. N. / The chemical characterization of calf brain microtubule protein subunits. In: Journal of Biological Chemistry. 1973 ; Vol. 248, No. 20. pp. 7253-7262.
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