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RGD peptides induce apoptosis by direct caspase-3 activation

Nature volume 397, pages 534–539 (1999)Cite this article

Abstract

Synthetic peptides containing the arginine–glycine–aspartate (RGD) motif have been used extensively as inhibitors of integrin–ligand interactions in studies of cell adhesion, migration, growth and differentiation1,2,3, because the RGD motif is an integrin-recognition motif found in many ligands. Here we report that RGD-containing peptides are able to directly induce apoptosis without any requirement for integrin-mediated cell clustering or signals. We show that RGD-containing peptides enter cells and directly induce autoprocessing and enzymatic activity of pro-caspase-3, a pro-apoptotic protein. Using the breast carcinoma cell line MCF-7, which has a functional deletion of the caspase-3 gene, we confirm that caspase-3 is required for RGD-mediated cell death. In addition to an RGD motif, pro-caspase-3 also contains a potential RGD-binding motif, aspartate–aspartate–methionine (DDM)4, near the site of processing to produce the p12 and p17 subunits5. On the basis of the ability of RGD–DDX interactions to trigger integrin activation6, we suggest that RGD peptides induce apoptosis by triggering conformational changes that promote pro-caspase-3 autoprocessing and activation. These findings provide an alternative molecular explanation for the potent pro-apoptotic properties of RGD peptides in models of angiogenesis, inflammation and cancer metastasis7,8,9.

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Figure 1: RGD-containing peptides induce lymphocyte apoptosis.
Figure 2: RGD peptides mediate their pro-apoptotic effects independently of integrin clustering and enter cells.
Figure 3: RGD-containing peptides induce pro-caspase-3 autoprocessing and activation directly in a cell-free system.
Figure 4: Caspase-3 is required for RGD-mediated cell death.
Figure 5: RGD peptides induce apoptosis and caspase-3 activation in lung fibroblasts cultured on type I collagen.

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Acknowledgements

We thank N. Shamsadeen, M. Zwede and D. Hardie for technical assistance. This work was funded by grants from the Arthritis Research Campaign (to D.P., D.S.-T., M.S.), The Royal Society (to J.M.L.), Cancer Research Campaign (to N.V.H.), Medical Research Council (to G.P., K.T., A.N.A.) and Wellcome Trust (C.D.B., D.L.S.).

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Author notes

  1. Christopher D. Buckley, Darrell Pilling and Nick V. Henriquez: These authors contributed equally to this work

Authors and Affiliations

  1. Division of Immunity and Infection, B15 2TT, Birmingham, UK

    Christopher D. Buckley, Darrell Pilling, Greg Parsonage, Katy Threlfall, Dagmar Scheel-Toellner, Janet M. Lord & Mike Salmon

  2. Institute for Cancer Studies, MRC Centre for Immune Regulation, The University of Birmingham, B15 2TT, Birmingham, UK

    Nick V. Henriquez

  3. Department of Neuroscience, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park, Third Avenue, Harlow, CM19 5AW, Essex, UK

    David L. Simmons

  4. Department of Clinical Immunology, Royal Free and University College Medical School, Pond Street, NW3 2PF, London, UK

    Arne N. Akbar

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Correspondence to Mike Salmon.

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Buckley, C., Pilling, D., Henriquez, N. et al. RGD peptides induce apoptosis by direct caspase-3 activation. Nature 397, 534–539 (1999). https://doi.org/10.1038/17409

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