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Mechanistic insight into taxol-induced cell death

Oncogene volume 27, pages 4580–4591 (2008)Cite this article

Abstract

We analysed the involvement of proteases during taxol-mediated cell death of human A549 non-small-cell lung carcinoma cells using a proteomics approach that specifically targets protein N termini and further detects newly formed N termini that are the result of protein processing. Our analysis revealed 27 protease-mediated cleavages, which we divided in sites C-terminal to aspartic acid (Asp) and sites C-terminal to non-Asp residues, as the result of caspase and non-caspase protease activities, respectively. Remarkably, some of the former were insensitive to potent pancaspase inhibitors, and we therefore suggest that previous inhibitor-based studies that report on the caspase-independent nature of taxol-induced cell death should be judged with care. Furthermore, many of the sites C-terminal to non-Asp residues were also uniquely observed in a model of cytotoxic granule-mediated cell death and/or found by in vitro cataloging human μ-calpain substrates using a similar proteomics technique. This thus raises the hypothesis that killing tumor cells by chemotherapy or by immune cells holds similar non-Asp-specific proteolytic components with strong indications to calpain activity.

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Abbreviations

AO:

acridine orange

CLC:

clathrin light chain

COFRADIC:

COmbined FRactional DIagonal Chromatography

KRT:

cytokeratin

LMP:

lysosomal membrane permeabilization

MAP1B:

microtubule-associated protein 1B

PCD:

programmed cell death

PTBP:

polypyrimidine tract-binding protein

SILAC:

stable isotope labeling by amino acids in cell culture

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Acknowledgements

FI is a Research Assistant of the Fund for Scientific Research—Flanders (Belgium). This work was supported by grants to JV and KG from the Inter University Attraction Poles (IAP-Phase VI), the Concerted Research Actions of the Ghent University and the Fund for Scientific Research—Flanders (Belgium). We thank Professor J Tavernier for providing the flow cytometry facilities.

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Authors and Affiliations

  1. Department of Medical Protein Research, VIB, Ghent, Belgium

    F Impens, P Van Damme, H Demol, J Van Damme, J Vandekerckhove & K Gevaert

  2. Department of Biochemistry, Ghent University, Ghent, Belgium

    F Impens, P Van Damme, H Demol, J Van Damme, J Vandekerckhove & K Gevaert

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  1. F Impens
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  2. P Van Damme
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Correspondence to K Gevaert.

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Impens, F., Van Damme, P., Demol, H. et al. Mechanistic insight into taxol-induced cell death. Oncogene 27, 4580–4591 (2008). https://doi.org/10.1038/onc.2008.96

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