3'-(p-Azidobenzamido)taxol photolabels the N-terminal 31 amino acids of β-tubulin

S. Rao, N. E. Krauss, J. M. Heerding, C. S. Swindell, I. Ringel, G. A. Orr, Susan Band Horwitz

Research output: Contribution to journalArticle

261 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)3132-3134
Number of pages3
JournalJournal of Biological Chemistry
Volume269
Issue number5
StatePublished - 1994

Fingerprint

Tubulin
Paclitaxel
Amino Acids
Microtubules
formic acid
Polymers
Binding Sites
Depolymerization
Protein Sequence Analysis
Bioactivity
Human Activities
Pharmaceutical Preparations
Antineoplastic Agents
Irradiation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Rao, S., Krauss, N. E., Heerding, J. M., Swindell, C. S., Ringel, I., Orr, G. A., & Band Horwitz, S. (1994). 3'-(p-Azidobenzamido)taxol photolabels the N-terminal 31 amino acids of β-tubulin. Journal of Biological Chemistry, 269(5), 3132-3134.

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 journalArticle

Rao, S, Krauss, NE, Heerding, JM, Swindell, CS, Ringel, I, Orr, GA & Band Horwitz, S 1994, '3'-(p-Azidobenzamido)taxol photolabels the N-terminal 31 amino acids of β-tubulin', Journal of Biological Chemistry, vol. 269, no. 5, pp. 3132-3134.
Rao S, Krauss NE, Heerding JM, Swindell CS, Ringel I, Orr GA et al. 3'-(p-Azidobenzamido)taxol photolabels the N-terminal 31 amino acids of β-tubulin. Journal of Biological Chemistry. 1994;269(5):3132-3134.
Rao, S. ; Krauss, N. E. ; Heerding, J. M. ; Swindell, C. S. ; Ringel, I. ; Orr, G. A. ; Band Horwitz, Susan. / 3'-(p-Azidobenzamido)taxol photolabels the N-terminal 31 amino acids of β-tubulin. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 5. pp. 3132-3134.
@article{0468d2a069ef4acb92177b2d96f7a3aa,
title = "3'-(p-Azidobenzamido)taxol photolabels the N-terminal 31 amino acids of β-tubulin",
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.",
author = "S. Rao and Krauss, {N. E.} and Heerding, {J. M.} and Swindell, {C. S.} and I. Ringel and Orr, {G. A.} and {Band Horwitz}, Susan",
year = "1994",
language = "English (US)",
volume = "269",
pages = "3132--3134",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "5",

}

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 -