Direct Photoaffinity Labeling of Tubulin With Taxol

Rao, Srinivasa P. S.; Horwitz, Susan Band; Ringel, Israel

doi:10.1093/jnci/84.10.785

Cited by 151 publications

(73 citation statements)

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“…tide-We decided to explore the potential of the direct photoaffinity labeling technique for providing information about the dolastatin 10-binding site on tubulin based on the successful studies with colchicine (17), paclitaxel (40), and GTP (9). Only in the latter case was a specific amino acid residue, Cys-12 of ␤-tubulin, identified, and Shivanna et al (9) proposed that exposure of tubulin to ultraviolet radiation resulted in generation of sulfhydryl free radicals as the reactive species.…”

Section: Direct Photoaffinity Labeling Of Tubulin By [ 3 H]dolastatinmentioning

confidence: 99%

Direct Photoaffinity Labeling by Dolastatin 10 of the Amino-terminal Peptide of β-Tubulin Containing Cysteine 12

Bai

Covell

Taylor

et al. 2004

Journal of Biological Chemistry

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The subunit protein of microtubules, the ␣␤-tubulin heterodimer, has a number of ligand-binding sites. These include the exchangeable and nonexchangeable GTP-binding sites and at least three reasonably well characterized sites that bind antimitotic drugs. The electron crystallographic model of the tubulin sheet polymer formed in the presence of zinc and paclitaxel and composed of antiparallel protofilaments has provided relatively detailed information about the locations of the nucleotide-binding sites and the paclitaxel site on the ␣␤-dimer (1).

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Section: Direct Photoaffinity Labeling Of Tubulin By [ 3 H]dolastatinmentioning

confidence: 99%

Direct Photoaffinity Labeling by Dolastatin 10 of the Amino-terminal Peptide of β-Tubulin Containing Cysteine 12

Bai

Covell

Taylor

et al. 2004

Journal of Biological Chemistry

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“…Assembly was monitored spectrophotometrically at 350 nm by following the change in turbidity that is representative of polymer mass (16). Electron microscopy was performed as described previously (3). Photoaffinity Labeling Studies and CNBr Digestion-For mapping studies, 1 mM GTP was added to 1 mg/ml MTP in assembly buffer at 37°C.…”

Section: Preparation and Polymerization Of Microtubule Protein (Mtp)-twomentioning

confidence: 99%

“…First, we have shown that direct photolabeling of tubulin with Taxol resulted in the labeling of ␤-tubulin (3). Further studies with [ 3 H]3Ј-(p-azidobenzamido)Taxol (where the arylazide was incorporated into the C-13 side chain) resulted in the isolation of a photolabeled peptide containing amino acid residues 1-31 in ␤-tubulin (10).…”

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confidence: 99%

Characterization of the Taxol Binding Site on the Microtubule

Rao¹,

He²,

Chakravarty³

et al. 1999

Journal of Biological Chemistry

169

103

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“…It has also shown some activity against various other tumour types (Etlinger, 1993;Forastiere, 1993;Einzig and Wiernik, 1995;Edelman and Gandara, 1996). Although the mechanism by which stable microtubules are induced by taxol are well established (Schiff et al, 1979;Rao et al, 1992Rao et al, , 1994, the molecular links between tubulin polymerisation, cell-cycle arrest, and ultimately cell death are not so well characterised. Given the broad spectrum of activity of taxol with regard to tumour type and genetic background, it is important that these mechanisms are resolved.…”

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confidence: 99%

Delayed expression of apoptosis in human lymphoma cells undergoing low-dose taxol-induced mitotic stress

2003

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The links between low-dose range taxol-induced mitotic arrest and the subsequent engagement of apoptosis are important for identifying the routes to therapeutic action. Here we have investigated the timing of cell-cycle perturbation and cell death responses following continuous exposure to clinically relevant drug concentrations (1 -20 nM). Following 8 h of exposure to taxol, the cell line DoHH2 (p53 wild type) exhibited mitotic arrest and engagement of apoptosis, whereas the cell line SU-DHL-4 (p53 mutant) breached cell-cycle arrest with progression to an abnormal cycle and a 24 h delay in the engagement of apoptosis. Imaging showed equivalent dysfunction of mitotic spindles in both cell lines. The results of kinetic analyses indicated that although cell death may occur at different stages of progression through mitosis and subsequent cell cycles, the overall kinetics of cell death relate to the rate of arrival at a critical event window in the cell cycle. We propose a simple model of low-dose taxol-induced cell death for cycling populations in which mitotic stress acts as a primary trigger for apoptosis with equivalent but potentially delayed outcomes. This view provides a rationale for the clinical effectiveness of this agent, independent of the initial capacity of the tumour cell to engage apoptosis due, for example, to mutant p53 expression. The results provide a perspective for the design of combination regimens that include low-dose taxol and a component that may disturb mitotic delivery.

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Direct Photoaffinity Labeling of Tubulin With Taxol

Abstract: This observation is the first step in a study to determine the binding site for taxol on the microtubule.

Cited by 151 publications

References 0 publications

Direct Photoaffinity Labeling by Dolastatin 10 of the Amino-terminal Peptide of β-Tubulin Containing Cysteine 12

Direct Photoaffinity Labeling by Dolastatin 10 of the Amino-terminal Peptide of β-Tubulin Containing Cysteine 12

Characterization of the Taxol Binding Site on the Microtubule

Delayed expression of apoptosis in human lymphoma cells undergoing low-dose taxol-induced mitotic stress

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