Bis(8-anilinonaphthalene-1-sulfonate) as a probe for tubulin decay

A. R S Prasad, Richard F. Luduena, Paul M. Horowitz

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

The fluorescent apolar probe bis(8-anilinonaphthalene-1-sulfonate) (Bis-ANS) has been used to detect structural correlates of the well-known but poorly understood decay of tubulin function, by which tubulin loses its ability to polymerize and bind drugs in a complex, time-dependent way. The present results indicate that the decay of tubulin is accompanied by the appearance of hydrophobic areas, which bind a total of six Bis-ANS molecules with a dissociation constant of 19 μM. This binding seems to be a result of localized structural changes that are taking place in the tubulin molecule and can be used as a probe for these changes. In particular, circular dichroism measurements revealed no significant changes in the average secondary structure of the protein during the time required for complete binding of the Bis-ANS molecules. Preincubation of tubulin with the antimitotic drugs colchicine, podophyllotoxin, and vinblastine slows the rate of appearance of the hydrophobic region. Vinblastine has the maximal effect followed by colchicine and podophyllotoxin. In contrast, preincubation with maytansine has no effect. In addition, lowering the temperature decreases the rate of appearance of this region. These results correlate with the effect of drugs on the alkylation of tubulin sulfhydryl groups by iodoacetamide [Luduena, R. F., & Roach, M. C. (1981) Biochemistry 20, 4444-4450] and with the ability of inhibitors of microtubule assembly to permit the polymerization of tubulin into nonmicrotubule structures.

Original languageEnglish (US)
Pages (from-to)739-742
Number of pages4
JournalBiochemistry
Volume25
Issue number3
StatePublished - 1986

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Tubulin
Podophyllotoxin
Vinblastine
Colchicine
Molecules
Maytansine
Iodoacetamide
Antimitotic Agents
Secondary Protein Structure
Biochemistry
Alkylation
Circular Dichroism
8-anilino-1-naphthalenesulfonic acid
Fluorescent Dyes
Microtubules
Polymerization
Pharmaceutical Preparations
Temperature
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Prasad, A. R. S., Luduena, R. F., & Horowitz, P. M. (1986). Bis(8-anilinonaphthalene-1-sulfonate) as a probe for tubulin decay. Biochemistry, 25(3), 739-742.

Bis(8-anilinonaphthalene-1-sulfonate) as a probe for tubulin decay. / Prasad, A. R S; Luduena, Richard F.; Horowitz, Paul M.

In: Biochemistry, Vol. 25, No. 3, 1986, p. 739-742.

Research output: Contribution to journalArticle

Prasad, ARS, Luduena, RF & Horowitz, PM 1986, 'Bis(8-anilinonaphthalene-1-sulfonate) as a probe for tubulin decay', Biochemistry, vol. 25, no. 3, pp. 739-742.
Prasad ARS, Luduena RF, Horowitz PM. Bis(8-anilinonaphthalene-1-sulfonate) as a probe for tubulin decay. Biochemistry. 1986;25(3):739-742.
Prasad, A. R S ; Luduena, Richard F. ; Horowitz, Paul M. / Bis(8-anilinonaphthalene-1-sulfonate) as a probe for tubulin decay. In: Biochemistry. 1986 ; Vol. 25, No. 3. pp. 739-742.
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