Abstract
Taxol possesses an unusual chemical structure, a unique mechanism of action, and demonstrated activity in human malignancies. It is the only antitumor agent that has a binding site on the microtubule polymer. The interaction of Taxol with the microtubule polymer results in the formation of stable bundles of cellular microtubules that are resistant to depolymerization. Although it has become evident that the microtubule, specifically β-tubulin, is the target for Taxol, no information is available on the binding site for the drug. In this report, we demonstrate that 3'-(p- azidobenzamido)taxol, an analogue with similar biological activities as Taxol, covalently binds to the N-terminal domain of β-tubulin after irradiation of the microtubule-drug complex. Taxol competes with [3H]3'-(p- azidobenzamido)-taxol binding, suggesting that the photoaffinity analog and Taxol are binding at the same or overlapping sites. Formic acid cleavage of [3H]3'-(p-azidobenzamido)-taxol-photolabeled β-tubulin and subsequent protein sequence and mass analyses have identified the N-terminal 31 amino acids as the major site for [3H]3'-(p-azidobenzamido)taxol photoincorporation.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3132-3134 |
| Number of pages | 3 |
| Journal | Journal of Biological Chemistry |
| Volume | 269 |
| Issue number | 5 |
| State | Published - 1994 |
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ASJC Scopus subject areas
- Biochemistry
Cite this
3'-(p-Azidobenzamido)taxol photolabels the N-terminal 31 amino acids of β-tubulin. / Rao, S.; Krauss, N. E.; Heerding, J. M.; Swindell, C. S.; Ringel, I.; Orr, G. A.; Band Horwitz, Susan.
In: Journal of Biological Chemistry, Vol. 269, No. 5, 1994, p. 3132-3134.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - 3'-(p-Azidobenzamido)taxol photolabels the N-terminal 31 amino acids of β-tubulin
AU - Rao, S.
AU - Krauss, N. E.
AU - Heerding, J. M.
AU - Swindell, C. S.
AU - Ringel, I.
AU - Orr, G. A.
AU - Band Horwitz, Susan
PY - 1994
Y1 - 1994
N2 - Taxol possesses an unusual chemical structure, a unique mechanism of action, and demonstrated activity in human malignancies. It is the only antitumor agent that has a binding site on the microtubule polymer. The interaction of Taxol with the microtubule polymer results in the formation of stable bundles of cellular microtubules that are resistant to depolymerization. Although it has become evident that the microtubule, specifically β-tubulin, is the target for Taxol, no information is available on the binding site for the drug. In this report, we demonstrate that 3'-(p- azidobenzamido)taxol, an analogue with similar biological activities as Taxol, covalently binds to the N-terminal domain of β-tubulin after irradiation of the microtubule-drug complex. Taxol competes with [3H]3'-(p- azidobenzamido)-taxol binding, suggesting that the photoaffinity analog and Taxol are binding at the same or overlapping sites. Formic acid cleavage of [3H]3'-(p-azidobenzamido)-taxol-photolabeled β-tubulin and subsequent protein sequence and mass analyses have identified the N-terminal 31 amino acids as the major site for [3H]3'-(p-azidobenzamido)taxol photoincorporation.
AB - Taxol possesses an unusual chemical structure, a unique mechanism of action, and demonstrated activity in human malignancies. It is the only antitumor agent that has a binding site on the microtubule polymer. The interaction of Taxol with the microtubule polymer results in the formation of stable bundles of cellular microtubules that are resistant to depolymerization. Although it has become evident that the microtubule, specifically β-tubulin, is the target for Taxol, no information is available on the binding site for the drug. In this report, we demonstrate that 3'-(p- azidobenzamido)taxol, an analogue with similar biological activities as Taxol, covalently binds to the N-terminal domain of β-tubulin after irradiation of the microtubule-drug complex. Taxol competes with [3H]3'-(p- azidobenzamido)-taxol binding, suggesting that the photoaffinity analog and Taxol are binding at the same or overlapping sites. Formic acid cleavage of [3H]3'-(p-azidobenzamido)-taxol-photolabeled β-tubulin and subsequent protein sequence and mass analyses have identified the N-terminal 31 amino acids as the major site for [3H]3'-(p-azidobenzamido)taxol photoincorporation.
UR - http://www.scopus.com/inward/record.url?scp=0028169689&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0028169689&partnerID=8YFLogxK
M3 - Article
C2 - 7906266
AN - SCOPUS:0028169689
VL - 269
SP - 3132
EP - 3134
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 5
ER -