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Functionalization of cobalt porphyrin–phospholipid bilayers with his-tagged ligands and antigens

Nature Chemistry volume 7, pages 438–446 (2015)Cite this article

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Abstract

Methods to attach polypeptides to lipid bilayers are often indirect and ineffective, and can represent a substantial bottleneck in the formation of functionalized lipid-based materials. Although the polyhistidine tag (his-tag) has been transformative in its simplicity and efficacy in binding to immobilized metals, the successful application of this approach has been challenging in physiological settings. Here we show that lipid bilayers containing porphyrin–phospholipid conjugates that are chelated with cobalt, but not with other metals, can effectively capture his-tagged proteins and peptides. The binding follows a Co(II) to Co(III) transition and occurs within the sheltered hydrophobic bilayer, resulting in an essentially irreversible attachment in serum or in a million fold excess of competing imidazole. Using this approach we anchored homing peptides into the bilayer of preformed and cargo-loaded liposomes to enable tumour targeting without disrupting the bilayer integrity. As a further demonstration, a synthetic protein fragment derived from the human immunodeficiency virus was bound to immunogenic liposomes for potent antibody generation for an otherwise non-antigenic peptide.

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Figure 1: His-tagged polypeptides bind and functionalize Co-PoP bilayers.
Figure 2: His-tagged protein binding to Co(III)-PoP liposomes.
Figure 3: Robust his-tagged protein binding to Co-PoP liposomes.
Figure 4: Binding of a short his-tagged RGD peptide to Co-PoP liposomes.
Figure 5: RGD-his targeting of cargo-loaded liposomes.
Figure 6: HIV peptide vaccination using immunogenic Co-PoP liposomes.

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Acknowledgements

The authors thank J. R. Morrow and R. B. Bankert for valuable discussions, A. Siegel for assistance with confocal microscopy and the NIH AIDS Reagent Program. This work was supported by grants from the National Institutes of Health (R01EB017270, DP5OD017898 and HL77258).

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Authors and Affiliations

  1. Department of Biomedical Engineering, University at Buffalo, State University of New York, Buffalo, 14260, New York, USA

    Shuai Shao, Jumin Geng & Jonathan F. Lovell

  2. Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, Buffalo, 14260, New York, USA

    Shuai Shao, Shobhit Gogia, Sriram Neelamegham & Jonathan F. Lovell

  3. Department of Microbiology and Immunology, University at Buffalo, State University of New York, Buffalo, 14260, New York, USA

    Hyun Ah Yi & Amy Jacobs

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Contributions

S.G. and S.N. designed and produced the his-tagged fluorescence protein reporter. S.S. performed the synthesis, chemical characterization, binding, targeting and immunization experiments. J.G. led the animal experiments. A.J. and H.A.Y. carried out the HIV entry inhibition experiments. S.S. and J.F.L. conceived the project, interpreted the data and wrote the manuscript.

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Correspondence to Jonathan F. Lovell.

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Shao, S., Geng, J., Ah Yi, H. et al. Functionalization of cobalt porphyrin–phospholipid bilayers with his-tagged ligands and antigens. Nature Chem 7, 438–446 (2015). https://doi.org/10.1038/nchem.2236

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