Abstract
As more and more studies utilize cell-penetrating peptides to deliver pharmacologically interesting substances, there is a growing need to understand their effect on the plasma membrane. If a cell-penetrating peptide together with its cargo is to be used as a drug, it is necessary to understand how the conjugate interacts with the plasma membrane to enter the cell. A key regulator of the transportation network in the cell is calcium. This chapter describes five methods that can be employed for understanding how the plasma membrane reacts to the presence of cell-penetrating peptides and the involvement of calcium.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Kaplan, I. M., Wadia, J. S., and Dowdy, S. F. (2005) Cationic TAT peptide transduction domain enters cells by macropinocytosis. J Control Release 102, 247–253.
Nakase, I., Tadokoro, A., Kawabata, N., Takeuchi, T., Katoh, H., Hiramoto, K., Negishi, M., Nomizu, M., Sugiura, Y., and Futaki, S. (2007) Interaction of arginine-rich peptides with membrane-associated proteoglycans is crucial for induction of actin organization and macropinocytosis. Biochemistry 46, 492–501.
Richard, J. P., Melikov, K., Brooks, H., Prevot, P., Lebleu, B., and Chernomordik, L. V. (2005) Cellular uptake of unconjugated TAT peptide involves clathrin-dependent endocytosis and heparan sulfate receptors. J Biol Chem 280, 15300–15306.
Duchardt, F., Fotin-Mleczek, M., Schwarz, H., Fischer, R., and Brock, R. (2007) A comprehensive model for the cellular uptake of cationic cell-penetrating peptides. Traffic 8, 848–866.
Padari, K., Säälik, P., Hansen, M., Koppel, K., Raid, R., Langel, Ü., and Pooga, M. (2005) Cell transduction pathways of transportans Bioconjug Chem 16, 1399–1410.
Fretz, M. M., Penning, N. A., Al-Taei, S., Futaki, S., Takeuchi, T., Nakase, I., Storm, G., and Jones, A. T. (2007) Temperature-, concentration- and cholesterol-dependent translocation of l- and d-octa-arginine across the plasma and nuclear membrane of CD34+ leukaemia cells. Biochem J 403, 335–342.
Tünnemann, G., Martin, R. M., Haupt, S., Patsch, C., Edenhofer, F., and Cardoso, M. C. (2006) Cargo-dependent mode of uptake and bioavailability of TAT-containing proteins and peptides in living cells. FASEB J 20, 1775–1784.
Bi, G. Q., Alderton, J. M., and Steinhardt, R. A. (1995) Calcium-regulated exocytosis is required for cell membrane resealing. J Cell Biol 131, 1747–1758.
Togo, T., Alderton, J. M., Bi, G. Q., and Steinhardt, R. A. (1999) The mechanism of facilitated cell membrane resealing. J Cell Sci 112, 719–731.
McNeil, P. L. and Steinhardt, R. A. (2003) Plasma membrane disruption: repair, prevention, adaptation. Annu Rev Cell Dev Biol 19, 697–731.
Miyake, K. and McNeil, P. L. (1995) Vesicle accumulation and exocytosis at sites of plasma membrane disruption. J Cell Biol 131, 1737–1745.
Yandek, L. E., Pokorny, A., Floren, A., Knoelke, K., Langel, Ü., and Almeida, P. F. (2007) Mechanism of the cell-penetrating peptide transportan 10 permeation of lipid bilayers. Biophys J 92, 2434–2444.
Howl, J., Jones, S., and Farquhar, M. (2003) Intracellular delivery of bioactive peptides to RBL-2H3 cells induces beta-hexosaminidase secretion and phospholipase D activation. Chembiochem 4, 1312–1316.
McNeil, P. L. (2002) Repairing a torn cell surface: make way, lysosomes to the rescue. J Cell Sci 115, 873–879.
Reddy, A., Caler, E. V., and Andrews, N. W. (2001) Plasma membrane repair is mediated by Ca(2+)-regulated exocytosis of lysosomes. Cell 106, 157–169.
Saar, K., Lindgren, M., Hansen, M., Eiriksdottir, E., Jiang, Y., Rosenthal-Aizman, K., Sassian, M., and Langel, Ü. (2005) Cell-penetrating peptides: a comparative membrane toxicity study. Anal Biochem 345, 55–65.
Johansson, H. J., El Andaloussi, S., Holm, T., Mäe, M., Jänes, J., Maimets, T., and Langel, Ü. (2008) Characterization of a novel cytotoxic cell-penetrating peptide derived from p14ARF protein. Mol Ther 16, 115–123.
Palm-Apergi, C., Lorents, A., Padari, K., Pooga, M., and Hällbrink, M. (2009) The membrane repair response masks membrane disturbances caused by cell-penetrating peptide uptake. FASEB J 23, 214–223.
Acknowledgments
This work was supported by the Jeansson foundation.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Palm-Apergi, C., Hällbrink, M. (2011). Calcium and Membrane Repair. In: Langel, Ü. (eds) Cell-Penetrating Peptides. Methods in Molecular Biology, vol 683. Humana Press. https://doi.org/10.1007/978-1-60761-919-2_11
Download citation
DOI: https://doi.org/10.1007/978-1-60761-919-2_11
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-60761-918-5
Online ISBN: 978-1-60761-919-2
eBook Packages: Springer Protocols