PMID: 2506927Jul 25, 1989Paper

Tubulin dimer dissociation detected by fluorescence anisotropy

Biochemistry
M R Mejillano, R H Himes

Abstract

We have demonstrated a concentration-dependent dissociation of bovine brain tubulin dimer covalently labeled with 5-[(4,6-dichlorotriazin-2-yl)amino]fluorescein (DTAF) or with fluorescein isothiocyanate (FITC) by fluorescence anisotropy and size-exclusion HPLC. The fluorescence anisotropy values decreased to a limiting value upon dilution of tubulin from 10(-5) to 8 x 10(-8) M. A dissociation constant in 0.1 M Pipes, pH 6.9, 1 mM EGTA, and 1 mM MgSO4 at 20 degrees C was estimated to be (8.4 x 10(-7) +/- (0.4 x 10(-7) M. Control experiments using monomeric and other dimeric proteins, urea-denatured tubulin, and DTAF-tubulin diluted into solutions of bovine serum albumin or unlabeled tubulin were consistent with the finding that the changes in anisotropy upon dilution are due to protein dissociation. These results were supported by size-exclusion HPLC data where an increase in the elution volume of DTAF-tubulin and FITC-tubulin was observed with decreasing protein concentrations. Reversibility of the dissociation process and the lack of denaturation at high dilution were shown by the ability of reconcentrated protein to assemble into microtubules to about the same extent as undiluted protein. Fluorescent lifetimes and limiting an...Continue Reading

References

Feb 24, 1976·Biochemistry·J D Shore, S K Chakrabarti
Sep 19, 1978·Biochemistry·H W Detrich, R C Williams
Jul 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·B Bhattacharyya, J Wolff
Mar 21, 1989·Biochemistry·D L SackettJ Wolff
Jun 13, 1988·Biochimica Et Biophysica Acta·J Algaier, R H Himes
Jul 1, 1988·Biochemical Pharmacology·W D SingerR H Himes
Sep 1, 1988·The Journal of Cell Biology·G P VigersJ R McIntosh
Mar 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·M L ShelanskiC R Cantor
May 26, 1972·Biochemical and Biophysical Research Communications·R J OwellenD W Donigian
Oct 1, 1983·The Journal of Cell Biology·P Wadsworth, R D Sloboda

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Citations

Sep 12, 2014·Journal of Fluorescence·Allan GuanK Scott Phillips
Jan 30, 1990·Biochemical and Biophysical Research Communications·M R MejillanoR H Himes
Apr 9, 1991·Biochemistry·D L Sackett, R E Lippoldt
Jun 1, 1990·Analytical Chemistry·H G Barth, B E Boyes
May 22, 2019·The Journal of Biological Chemistry·Felipe Montecinos-FranjolaDan L Sackett
Mar 5, 2016·The Journal of Biological Chemistry·Felipe Montecinos-FranjolaDan L Sackett
Mar 1, 1992·European Journal of Biochemistry·D Panda, B Bhattacharyya
Jul 14, 1995·The Journal of Biological Chemistry·J Wolff, L Knipling

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