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Heat-shock protein 60 translocates to the surface of apoptotic cells and differentiated megakaryocytes and stimulates phagocytosis

Cellular and Molecular Life Sciences volume 68, pages 1581–1592 (2011)Cite this article

Abstract

Heat-shock protein 60 (Hsp60) is a highly conserved stress protein which has chaperone functions in prokaryotes and mammalian cells. Hsp60 is associated with the mitochondria and the plasma membrane through phosphorylation by protein kinase A, and is incorporated into lipid membranes as a protein-folding chaperone. Its diverse intracellular chaperone functions include the secretion of proteins where it maintains the conformation of precursors and facilitates their translocation through the plasma membrane. We report here that Hsp60 is concentrated in apoptotic membrane blebs and translocates to the surface of cells undergoing apoptosis. Hsp60 is also enriched in platelets derived from terminally differentiated megakaryocytes and expressed at the surface of senescent platelets. Furthermore, the exposure of monocytic U937 cells to Hsp60 enhanced their phagocytic activity. Our results suggests that externalized Hsp60 in apoptotic cells and senescent platelets influences events subsequent to apoptosis, such as the clearance of apoptotic cells by phagocytes.

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Acknowledgments

This work was funded by the National Medical Research Council (NMRC) of Singapore and the Singapore Totalisator Board. We thank Dr. Yaw Chyn Lim, Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, for her critical appraisal of sections of our work.

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Affiliations

  1. Department of Surgery, Singapore General Hospital, Singapore, Singapore

    Yaw Chong Goh

  2. Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore

    Celestial T. Yap, Bao Hua Huang & Bernard P. Leung

  3. Institute of Structural and Molecular Biology, University of Edinburgh, Edinburgh, UK

    Andrew D. Cronshaw

  4. Department of Surgery, Chinese University of Hong Kong, Shatin, Hong Kong

    Paul B. S. Lai

  5. Department of Respiratory Medicine, University of Hull, Hull, UK

    Simon P. Hart

  6. MRC Centre for Inflammation Research, Queen’s Medical Research Institute, University of Edinburgh, Edinburgh, UK

    Ian Dransfield

  7. Tissue Injury and Repair Group, MRC Centre for Regenerative Medicine, University of Edinburgh, 49 Little France Crescent, Edinburgh, EH16 4SB, UK

    James A. Ross

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  1. Yaw Chong Goh
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Correspondence to Yaw Chong Goh or James A. Ross.

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Yaw Chong Goh and Celestial T. Yap are co-authors, and Yaw Chong Goh and James A. Ross are co-corresponding authors.

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Mass spectrometry analysis identifies Hsp60 as the antigen recognised by BOB78 antibody. Protein bands immunoprecipitated with BOB78 antibody were digested with trypsin, and peptides from the digestions were examined by LC-MS and MALDI-MS mass spectrometers. The samples were analysed on a Voyager DE-STR MALDI-TOF MS (Applied Biosystems) and the processed spectra searched against the NCBI non-redundant database such as SwissProt using Protein Prospector.

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Goh, Y.C., Yap, C.T., Huang, B.H. et al. Heat-shock protein 60 translocates to the surface of apoptotic cells and differentiated megakaryocytes and stimulates phagocytosis. Cell. Mol. Life Sci. 68, 1581–1592 (2011). https://doi.org/10.1007/s00018-010-0534-0

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  • DOI: https://doi.org/10.1007/s00018-010-0534-0

Keywords

  • Hsp60
  • Apoptosis
  • Translocation
  • Platelets
  • Phagocytosis