Abstract
In the study of cell-penetrating and membrane-translocating peptides, a fundamental question occurs as to the contribution arising from fundamental peptide–membrane interactions, relative to the contribution arising from the biology and energy of the cell, mostly occurring in the form of endocytosis and subsequent events. A commonly used approach to begin addressing these mechanistic questions is to measure the degree to which peptides can interact with, and physically disrupt, the integrity of synthetic lipid bilayers. Here, we describe a set of experimental methods that can be used to measure the potency, kinetics, transience, and the effective size of peptide-induced membrane disruption.
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References
Montrose K, Yang Y, Sun X, Wiles S, Krissansen GW (2013) Xentry, a new class of cell-penetrating peptide uniquely equipped for delivery of drugs. Sci Rep 3:1661
Dupont E, Prochiantz A, Joliot A (2011) Penetratin story: an overview. Methods Mol Biol 683:21–29
Schmidt N, Mishra A, Lai GH, Wong GC (2010) Arginine-rich cell-penetrating peptides. FEBS Lett 584:1806–1813
Chugh A, Eudes F, Shim YS (2010) Cell-penetrating peptides: nanocarrier for macromolecule delivery in living cells. IUBMB Life 62:183–193
Said HF, Saleh AF, Abes R, Gait MJ, Lebleu B (2010) Cell penetrating peptides: overview and applications to the delivery of oligonucleotides. Cell Mol Life Sci 67:715–726
Heitz F, Morris MC, Divita G (2009) Twenty years of cell-penetrating peptides: from molecular mechanisms to therapeutics. Br J Pharmacol 157:195–206
He J, Kauffman WB, Fuselier T, Naveen SK, Voss TG, Hristova K, Wimley WC (2013) Direct cytosolic delivery of polar cargo to cells by spontaneous membrane-translocating peptides. J Biol Chem 288:29974–29986
He J, Hristova K, Wimley WC (2012) A highly charged voltage sensor helix translocates spontaneously across membranes. Angew Chem Int Ed 51:7150–7153
Marks JR, Placone J, Hristova K, Wimley WC (2011) Spontaneous membrane-translocating peptides by orthogonal high-throughput screening. J Am Chem Soc 133:8995–9004
Ladokhin AS, Wimley WC, Hristova K, White SH (1997) Mechanism of leakage of contents of membrane vesicles determined by fluorescence requenching. Methods Enzymol 278:474–486
Ladokhin AS, Wimley WC, White SH (1995) Leakage of membrane vesicle contents: determination of mechanism using fluorescence requenching. Biophys J 69:1964–1971
Rausch JM, Wimley WC (2001) A high-throughput screen for identifying transmembrane pore-forming peptides. Anal Biochem 293:258–263
Krauson AJ, He J, Wimley WC (2012) Determining the mechanism of membrane permeabilizing peptides: identification of potent, equilibrium pore-formers. Biochim Biophys Acta 1818:1625–1632
Wiedman G, Fuselier T, He J, Searson PC, Hristova K, Wimley WC (2014) Highly efficient macromolecule-sized poration of lipid bilayers by a synthetically evolved peptide. J Am Chem Soc 136:4724–4731
Wimley WC (2010) Describing the mechanism of antimicrobial peptide action with the interfacial activity model. ACS Chem Biol 5:905–917
Wimley WC, Hristova K (2011) Antimicrobial peptides: successes, challenges and unanswered questions. J Membr Biol 239:27–34
Bechinger B, Lohner K (2006) Detergent-like actions of linear amphipathic cationic antimicrobial peptides. Biochim Biophys Acta 1758:1529–1539
Shai Y, Oren Z (2001) From “carpet” mechanism to de-novo designed diastereomeric cell-selective antimicrobial peptides. Peptides 22:1629–1641
Ladokhin AS, White SH (2001) ‘Detergent-like’ permeabilization of anionic lipid vesicles by melittin. Biochim Biophys Acta 1514:253–260
Wiedman G, Herman K, Searson P, Wimley WC, Hristova K (2013) The electrical response of bilayers to the bee venom toxin melittin: evidence for transient bilayer permeabilization. Biochim Biophys Acta 1828:1357–1364
Shai Y (2002) Mode of action of membrane active antimicrobial peptides. Biopolymers 66:236–248
Sengupta D, Leontiadou H, Mark AE, Marrink SJ (2008) Toroidal pores formed by antimicrobial peptides show significant disorder. Biochim Biophys Acta 1778:2308–2317
Ludtke SJ, He K, Heller WT, Harroun TA, Yang L, Huang HW (1996) Membrane pores induced by magainin. Biochemistry 35:13723–13728
Yang L, Harroun TA, Weiss TM, Ding L, Huang HW (2001) Barrel-stave model or toroidal model? A case study on melittin pores. Biophys J 81:1475–1485
White SH, Wimley WC, Ladokhin AS, Hristova K (1998) Protein folding in membranes: determining the energetics of peptide-bilayer interactions. Methods Enzymol 295:62–87
Ladokhin AS, Jayasinghe S, White SH (2000) How to measure and analyze tryptophan fluorescence in membranes properly, and why bother? Anal Biochem 285:235–245
Parente RA, Nir S, Szoka F (1990) Mechanism of leakage of phospholipid vesicle contents induced by the peptide GALA. Biochemistry 29:8720–8728
Krauson AJ, He J, Wimley WC (2012) Gain-of-function analogues of the pore-forming peptide melittin selected by orthogonal high-throughput screening. J Am Chem Soc 134:12732–12741
Hristova K, Selsted ME, White SH (1997) Critical role of lipid composition in membrane permeabilization by rabbit neutrophil defensins. J Biol Chem 272:24224–24233
Goñi FM, Ostolaza H (1998) E. coli a-hemolysin: a membrane-active protein toxin. Braz J Med Biol Res 31:1019–1034
Ladokhin AS, Selsted ME, White SH (1997) Sizing membrane pores in lipid vesicles by leakage of co-encapsulated markers: pore formation by melittin. Biophys J 72:1762–1766
Nayar R, Hope MJ, Cullis PR (1989) Generation of large unilamellar vesicles from long-chain saturated phosphatidylcholines by extrusion technique. Biochim Biophys Acta 986:200–206
Bartlett GR (1959) Phosphorus assay in column chromatography. J Biol Chem 234:466–468
Stewart JC (1980) Colorimetric determination of phospholipids with ammonium ferrothiocyanate. Anal Biochem 104:10–14
Hristova K, Kenworthy AK, McIntosh TJ (1995) Effect of bilayer composition on the phase behavior of liposomal suspensions containing poly(ethylene glycol)-lipids. Macromolecules 28:7693–7699
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Wimley, W.C. (2015). Determining the Effects of Membrane-Interacting Peptides on Membrane Integrity. In: Langel, Ü. (eds) Cell-Penetrating Peptides. Methods in Molecular Biology, vol 1324. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2806-4_6
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DOI: https://doi.org/10.1007/978-1-4939-2806-4_6
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2805-7
Online ISBN: 978-1-4939-2806-4
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