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Bacterial Hsp70 (DnaK) and mammalian Hsp70 interact differently with lipid membranes

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Cell Stress and Chaperones Aims and scope

Abstract

The cellular response to stress is orchestrated by the expression of a family of proteins termed heat shock proteins (hsp) that are involved in the stabilization of basic cellular processes to preserve cell viability and homeostasis. The bulk of hsp function occurs within the cytosol and subcellular compartments. However, some hsp have also been found outside cells released by an active mechanism independent of cell death. Extracellular hsp act as signaling molecules directed at activating a systemic response to stress. The export of hsp requires the translocation from the cytosol into the extracellular milieu across the plasma membrane. We have proposed that membrane insertion is the initial step in this export process. We investigated the interaction of the major inducible hsp from mammalian (Hsp70) and bacterial (DnaK) species with liposomes. We found that mammalian Hsp70 displayed a high specificity for negatively charged phospholipids, such as phosphatidyl serine, whereas DnaK interacted with all lipids tested regardless of the charge. Both proteins were inserted into the lipid bilayer as demonstrated by resistance to acid or basic washes that was confirmed by partial protection from proteolytic cleavage. Several regions of mammalian Hsp70 were inserted into the membrane with a small portion of the N-terminus end exposed to the outer phase of the liposome. In contrast, the N-terminus end of DnaK was inserted into the membrane, exposing the C-terminus end outside the liposome. Mammalian Hsp70 was found to make high oligomeric complexes upon insertion into the membranes whereas DnaK only formed dimers within the lipid bilayer. These observations suggest that both Hsp70s interact with lipids, but mammalian Hsp70 displays a high degree of specificity and structure as compared with the bacterial form.

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Acknowledgments

This work was supported by the National Institutes of Health, grant numbers GM R01 09845 and GM R25 083275.

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Authors and Affiliations

  1. Initiative for Maximizing Student Development (IMSD) Program, La Jolla, CA, USA

    Victor Lopez

  2. Department of Surgery, School of Medicine, University of California San Diego, La Jolla, CA, USA

    David M. Cauvi & Antonio De Maio

  3. Uniformed Services University, Bethesda, MD, USA

    Nelson Arispe

  4. Center for Investigations of Health and Education Disparities, La Jolla, CA, USA

    Antonio De Maio

  5. Department of Neuroscience, School of Medicine, University of California San Diego, La Jolla, CA, USA

    Antonio De Maio

  6. University of California San Diego, 9500 Gilman Drive, #0739, La Jolla, CA, 92093-0739, USA

    Antonio De Maio

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  1. Victor Lopez
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  2. David M. Cauvi
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  3. Nelson Arispe
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  4. Antonio De Maio
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Correspondence to Antonio De Maio.

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Lopez, V., Cauvi, D.M., Arispe, N. et al. Bacterial Hsp70 (DnaK) and mammalian Hsp70 interact differently with lipid membranes. Cell Stress and Chaperones 21, 609–616 (2016). https://doi.org/10.1007/s12192-016-0685-5

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  • DOI: https://doi.org/10.1007/s12192-016-0685-5

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